G-Biosciences Molecular Biology Handbook & Selection Guide • Protein Estimation Assays • Lysis Buffers & Systems • Apoptosis Assays • Protein Fractionation Kits • Cytotoxicity Assays • Dialysis (Micro) System • SAM Methyltransferase Assays • Electrophoresis Clean-Up • Protease Assays • Concentration Systems • Phosphatase Assays • Contamination Removal • Peroxide Assay

• Protease Inhibitor Cocktails • Proteomic Grade Detergents • Individual Protease Inhibitors • Research Grade Detergents • Protease Assays • Non-Ionic, Ionic & Zwitterionic • Proteases for Mass Spec. • Detergent Estimations • Sequencing Grade Proteases • Detergent Removal Systems

• 1-Hour Western System • Gel Preparation Chemicals • Transfer Buffers & Membranes • Protein Marker Ladders • Membrane Stains • Electrophoresis Buffers • Blocking Buffers • Reducing & Alkylating Reagents • Secondary Antibodies • Protein Gel Stains • Detection Reagents • Reprobing Reagents

• Protein Sample Preparation • Affinity Resins • Protein Clean-Up Systems • 6X His Protein Purification Kits • Electrophoresis Reagents • GST Protein Purification Kits • Mass Spec Grade Protease • Antibody Purification • InGel Digestion Kits • Activated Resins • Peptide Generation Reagents • Buffers & Reagents

• Biotin Labeling • Carrier Proteins • Cell Surface Protein Labeling • Peptide Coupling Systems • Agarose Coupling Kits • Antibody Purification Resins • Fluorescent Dye Labeling Kits • Antibody Fragmentation Kits • Enzyme Labeling Systems

• Coated Plates • Blocking Buffers • Homobifunctional • Wash Buffers • Heterobifunctional • Secondary Antibodies • Optimizer Systems • Detection Reagents • Cross-Linking Systems • Antibody Labeling Systems

• Apoptosis Assays • DNA Isolation • Cytotoxicity Assays • Transformation & Screening • SAM Methyltransferase Assays • Polymerase Chain Reaction • Protease Assays • Agarose Electrophoresis • Phosphatase Assays • RNA Isolation • Peroxide Assay • Yeast Transformation • ELISA Table Of Contents Genomic DNA Purification 2 PCR Clean Up...... 15 Spin-OUT™ for PCR...... 15 Introduction...... 2 Reverse Transcriptases (RT)...... 15 Purest DNA with OmniPrep™...... 2 ™ M-MLV Reverse Transcriptase...... 15 OmniPrep ...... 2 AMV Reverse Transcriptase...... 15 OmniPrep™ for Tissue...... 2 OmniPrep™ for Blood...... 3 RT-PCR Kits...... 16 OmniPrep™ for Fungi...... 3 First-strand cDNA Synthesis Kit...... 16 OmniPrep™ for Gram Positive Bacteria...... 3 1-Step RT-PCR Kit...... 16 ™ 2-Step RT-PCR Kit...... 16 OmniPrep for Yeast...... 3 ™ OmniPrep™ for Plant...... 4 Tri-Xtract ...... 16 OmniPrep™ for Mouse Tail...... 4 RNA Purification 17 ™ Spin Format Isolation with GET ...... 4 Arrest™ Extraction Buffer...... 17 ™ GET DNA Template...... 4 GET™ Total RNA...... 17 ™ GET Plant DNA Template...... 4 TOTAL Arrest™ ...... 17 Protein Free DNA with XIT™...... 5 NUCLEIC dotMETRIC™ Assay...... 18 XIT™ DNA from Tissue...... 5 RNase-DETECT™ ...... 18 XIT™ DNA from Cells...... 5 RNaseOUT™ ...... 18 XIT™ DNA from Blood...... 5 RNasin ...... 18 XIT™ DNA from Buccal Cells...... 6 XIT™ DNA from FFPE Tissue...... 6 Nucleic Acid Electrophoresis 19 XIT™ DNA from Mouse Tail...... 6 Geno-ElectroPhore™...... 19 XIT™ DNA from Plant Tissues...... 6 Power Supplies...... 19 XIT™ DNA from Gram Positive Bacteria...... 7 GT 250 Power Pack™...... 19 XIT™ DNA from Gram Negative Bacteria...... 7 E-Power™ 150 Power Supply...... 19 XIT™ DNA from Yeast...... 7 E-Power™ 100 Power Supply...... 19 XIT™ Mitochondrial DNA...... 7 G-CAPSULE™...... 20 PCR Template DNA...... 8 GET™ AGAROSE DNA...... 20 OmniTemplate™...... 8 geneEXIT™...... 20 Rapid DNA Template Prep™...... 8 Glow™ Dyes ...... 21 Yeast Geno-DNA-Template™...... 8 DNA Ladders...... 22 Plant Geno-DNA-Template™...... 9 DNAmark™ Logic DNA Ladder...... 22 High Molecular Weight DNA...... 9 DNAmark™ 100bp Plus Ladder...... 22 MegaLong™...... 9 DNAmark™ 100bp Ladder...... 22 Plasmid DNA Tools 10 Accessories, Buffers & Chemicals...... 23 Electrophoresis Running Buffers...... 23 Plasmid Isolation...... 10 Agarose Powders...... 23 GET™ Plasmid DNA Miniprep...... 10 LabSafe™ Nucleic Acid Stain...... 23 GET™ Plasmid DNA Mediprep...... 10 FASTsilver™ ...... 23 GET™ Plasmid DNA Maxiprep...... 10 Bacteria Transformation...... 11 Hybridization Buffers 24 ™ Z-Competent™ E. coli Transformation...... 11 EKONO ...... 24 ™ Plasmid Screening Reagents...... 11 HYB-LINK ...... 24 Plasmid Screening...... 11 Other Hybridization Buffers...... 24 Plasmid Screening ToothPick™...... 11 Yeast Research Tools 25 ToothPick™-PCR...... 11 EZ Yeast™ Plasmid Prep...... 25 DNA Cleanup & Concentration 12 Fast Yeast Transformation™...... 25 Yeast Related Buffers...... 25 GET™ CLEAN DNA...... 12 Yeast Geno-DNA-Template™...... 26 pinkCLEANUP™ ...... 12 LongLife™ Zymolyase®...... 26 Genomic Tube-O-DIALYZER™...... 12 LongLife™ Enzyme Preparations...... 26 Spin-OUT™ for PCR...... 12 DNA OUT™...... 27 Spin-OUT™...... 13 GET™ AGAROSE DNA...... 13 Molecular Biology Buffers & Chemicals...... 27 geneEXIT™...... 13 Molecular Biology Universal Kit...... 27 Research Grade Water...... 27 PCR/ RT-PCR 14 Endotoxin Free Water...... 27 DNA Polymerases...... 14 Molecular Grade Water...... 27 Taq Polymerase...... 14 DEPC-Treated Water...... 27 Taq Polymerase (2X MasterMix)...... 14 Proteomic Grade Water...... 27 Pfu Polymerase...... 14 Buffers & Reagents...... 28 Pfu Polymerase (2X MasterMix)...... 14 Deoxynucleotides...... 15 Deoxynucleotides (dNTPs) Set...... 15 Deoxynucleotides (dNTPs) Mix...... 15 For further details, visit VWR.com 1 Genomic DNA Purification Introduction FEATURES ———————————————————————————— • High Yield, ~100kb genomic DNA A selection of genomic isolation kits are offered that purify high • A260/A280 between 1.8 and 2.0 quality genomic DNA from a wide variety of sources and for a wide • Adaptable for variety of samples array of applications. • Simple two tube method, 20-40 minutes OmniPrep™ genomic DNA kits are for ultra pure genomic DNA • No toxic chemicals, no phenol, no hazardous waste that is suitable for all downstream applications. These kits are fully • Suitable for ~150 preparations depending on sample scalable for large genomic DNA isolations. The procedure is slightly APPLICATIONS more involved to ensure ultra pure DNA. • Extraction of pure genomic DNA from cells and tissues, plant, GET™ genomic DNA kits are spin column format kits for the rapid fungi, bacteria, whole blood, and other samples isolation of genomic DNA from small sample sizes. • Extraction of high quality, 100kb genomic DNA ™ XIT DNA kits produce protein free, high quality DNA and use CITED REFERENCES protein digestion and precipitation followed by genomic DNA Staley, C.A. et al (2012) Gene. 496:118 purification. No chloroform or phenol extraction required. Li, Z. et al (2010) Protein Expression and Purification. 72:113 OmniTemplate™ genomic DNA kits are for the rapid generation of Li, Z. et al (2010) Biochem and Biophys Res Comm. 402:519 Li, Z. et al (2009) Protein Expression and Purification. 67:175 PCR ready genomic templates. Lin-Cereghino, J. et al (2008) Yeast. 25:293 ™ MegaLong isolates high molecular weight genomic DNA from a Choi, Y. and Shim, W. (2008) Microbiology. 154:326 variety of samples, including animal tissues, cultured cells, whole Choi, Y. et al (2008) Mycologia. 100:701 blood, bacterial and yeast. Whitaker, V. et al (2007) J. Amer. Soc. Hort. Sci. 132:534 Lee, B et al (2005) Genome. 48:1104 ™ Pliss, L. et al (2004) J. Neurochem. 91:1082 Purest DNA with OmniPrep Jacobs-Helber, S. et al (2002) JBC. 277:4859 Li, X. et al (2002) Genome. 45:229 ———————————————————————————— Villar, M. et al (2001) J. Bact. 183:55 ™ OmniPrep Cat. No. Description Size ———————————————————————————— 82021-538 OmniPrep™ >150 Preps High quality genomic DNA from diverse sources Isolates high quality genomic DNA from many different species and tissue types including animal, plant, bacteria, yeast, fungi, whole ™ blood, and cells in culture. DNA can be isolated from samples high OmniPrep for Tissue in polysaccharides or other contaminants that are difficult to remove ———————————————————————————— from the DNA preparations. High quality genomic DNA from animal tissue Based on rapid precipitation technique that uses unique OmniPrep™ for Tissue kit has been modified to specifically isolate precipitation reagents to isolate genomic DNA free from proteins and high quality genomic DNA from tissue samples, including fresh, RNA. Pure genomic DNA is isolated in 20-40 minutes, depending frozen, fixed or paraffin-embedded tissue. The kit isolates DNA from on the tissue sample type used. The resulting genomic DNA is bodily fluids, including plasma, serum, amniotic fluid, semen and CSF. visualized on an agarose gel as a large single band, demonstrating Also isolates high quality DNA from cultured cells, non-mammalian high quality and minimal shearing. nucleated blood and gram-negative bacteria.

The kit isolates high purity (A260/A280 ratios of 1.8 to 2) DNA (~100kbp) and the yield is 0.5-10μg/mg tissue, 25-50μg/ml gram negative bacteria culture and 0.1-40μg/ml body fluid, dependent on starting material and quantity. If used according to the protocols, the kit purifies DNA from a total of 2gm solid tissue, 1x109 cultured cells and 1x1010 gram-negative bacteria. The kit uses a rapid precipitation technique that uses unique precipitation reagents to isolate genomic DNA free from proteins and RNA. Pure genomic DNA is isolated in 20-40 minutes, depending on the tissue type used. Suitable for: • Mammalian tissue (fresh or frozen) Figure 1: Omniprep™ scheme. • Cultured cells Suitable for a wide range of tissues and samples, including: • Ethanol or formalin fixed tissue • Mammalian tissues (fresh or frozen) • Paraffin embedded tissue • Cultured cells • Bodily fluids, i.e. plasma, sera, amniotic fluid, semen & CSF • Paraffin embedded tissues • Nucleated blood cells from bird, fish and frog • Ethanol or formalin fixed tissues • Gram negative bacteria • Nucleated blood cells from bird, fish and frog FEATURES • Bacteria (Gram positive and negative) • High Yield, ~100kb genomic DNA • Plant tissues (fresh or frozen, rich in polysaccharides) • A /A between 1.8 and 2.0 • Mouse tail 260 280 • Simple two tube method, 20-40 minutes • Yeast • No toxic chemicals, no phenol, no hazardous waste • Fungi • Blood APPLICATIONS • Blood stained and bodily fluid stained material • Extraction of pure genomic DNA from mammalian tissues • Soil • Suitable for fresh, fixed, frozen or paraffin embedded tissue The OmniPrep™ kit contains individual protocols for each different Cat. No. Description Size sample type and instructions on how to adapt for large sample 95029-204 OmniPrep™ for Tissue For ~2gm tissue volumes. 2 For further details, visit VWR.com Genomic DNA Purification OmniPrep™ for Blood OmniPrep™ for Gram Positive Bacteria ———————————————————————————— ———————————————————————————— High quality genomic DNA from blood High quality genomic DNA from bacteria OmniPrep™ for Blood kit isolates high quality genomic DNA from OmniPrep™ for Gram Positive Bacteria kit isolates high quality blood samples, including whole blood, buffy coats, packed cells and genomic DNA from Gram-positive bacteria. The kit isolates high ™ bone marrow. OmniPrep for Blood isolates high purity (A260/A280 purity (A260/A280 ratios of 1.8 to 2.0) DNA (~100kbp) and the yield is ratios of 1.8 to 2.0) DNA (~100kbp) and the yield is between 5-30µg/ 25-50μg/ml Gram-positive culture. If used according to the protocols ml, dependent on starting material and quantity. This kit is suitable this kit purifies DNA from 100ml Gram-positive bacteria culture. for processing up to 100ml blood. The kit uses a rapid precipitation technique that uses unique The kit uses a rapid precipitation technique that uses unique precipitation reagents to isolate genomic DNA free from proteins and precipitation reagents to isolate genomic DNA free from proteins and RNA. This kit is supplied with additional LongLife™ Lysozyme for rapid RNA. Pure genomic DNA is isolated in 20-40 minutes, depending on digestion of bacteria cell walls and DNA release. Pure genomic DNA the sample type used. OmniPrep™ for Blood isolates genomic DNA is isolated in 20-40 minutes, depending on the bacteria cell type from: used. • Whole blood FEATURES • Buffy Coats • High Yield, ~100kb genomic DNA • Packed cells • A260/A280 between 1.8 and 2.0 • Bone marrow • Simple two tube method, 20-40 minutes FEATURES • Supplied with LongLife™ Lysozyme for rapid bacterial digestion • High Yield, ~100kb genomic DNA • No toxic chemicals, no phenol, no hazardous waste

• A260/A280 between 1.8 and 2.0 APPLICATIONS • Simple two tube method, 20-40 minutes • Extraction of pure genomic DNA from Gram negative bacteria • No toxic chemicals, no phenol, no hazardous waste APPLICATIONS Cat. No. Description Size ™ • Extraction of pure genomic DNA from blood 95029-210 OmniPrep for Gram Positive Bacteria For 100ml culture • Suitable for whole blood, buffy coats, packed cells & bone marrow • Extraction of high quality, 100kb genomic DNA Cat. No. Description Size OmniPrep™ for Yeast 95029-206 OmniPrep™ for Blood For 100ml blood ———————————————————————————— High quality genomic DNA from yeast OmniPrep™ for Yeast kit isolates high quality genomic DNA from

™ yeast. The kit isolates high purity (A260/A280 ratios of 1.8 to 2.0) DNA OmniPrep for Fungi (~100kbp) and the yield is 3-6μg/ml yeast culture. If used according ———————————————————————————— to the protocols this kit purifies DNA from 300ml yeast culture. High quality genomic DNA from fungal tissue The kit uses a rapid precipitation technique that uses unique OmniPrep™ for Fungi kit isolates high quality genomic DNA from precipitation reagents to isolate genomic DNA free from proteins and fungal samples. The kit isolates high purity (A260/A280 ratios of 1.8 to RNA. This kit is supplied with a proprietary Yeast Suspension Buffer 2.0) DNA (~100kbp) and the yield is 0.2-1µg/5mg fungal samples. and LongLife™ Zymolyase® for the rapid resuspension and digestion If used according to the protocols this kit purifies DNA from 1-2gm of the yeast cell walls for DNA release. Pure genomic DNA is isolated fungal tissues. in 20-40 minutes, depending on the tissue type used. Based on a rapid precipitation technique that uses unique FEATURES precipitation reagents to isolate genomic DNA free from proteins and • High Yield, ~100kb genomic DNA RNA. ™ • A260/A280 between 1.8 and 2.0 This kit is supplied with Molecular Grinding Resin for the rapid • Simple two tube method, 20-40 minutes release of DNA from the fungal tissue. Pure genomic DNA is isolated • Supplied with Yeast Suspension Buffer and LongLife™ Zymolyase® in 20-40 minutes, depending on the tissue type used. for efficient DNA release FEATURES • No toxic chemicals, no phenol, no hazardous waste • High Yield, ~100kb genomic DNA APPLICATIONS • A /A between 1.8 and 2.0 260 280 • Extraction of pure genomic DNA from yeast tissue • Simple two tube method, 20-40 minutes • Extraction of high quality, 100kb genomic DNA • Molecular Grinding Resin™ for efficient fungal disruption • No toxic chemicals, no phenol, no hazardous waste Cat. No. Description Size APPLICATIONS 95029-214 OmniPrep™ for Yeast For 300ml culture • Extraction of pure genomic DNA from fungal tissue CITED REFERENCES Lorch, J. et al (2010) J Vet Diagn Invest. 22:224 Szabo, L. J. (2007) Mol. Ecol. Notes. 7:92 Ordonez, M. E. and Kolmer, J. A. (2007) Phytopathology. 97:574 Sagaram, U. S. et al (2006) Mol. Plant Pathol. 7:381 Mertens, J. A. et al (2006) Arch. Microbiol. 186:41

Cat. No. Description Size 95029-212 OmniPrep™ for Fungi For 2gm fungi

For further details, visit VWR.com 3 Genomic DNA Purification OmniPrep™ for Plant Spin Format Isolation with GET™ ———————————————————————————— ———————————————————————————— High quality genomic DNA from plant ™ OmniPrep™ for Plant kit isolates high quality genomic DNA from GET DNA Template plant samples, including fresh and frozen plant tissues. The kit ———————————————————————————— Spin column genomic DNA isolation isolates high purity (A260/A280 ratios of 1.8 to 2.0) DNA (~100kbp) and the yield is 0.5-3μg/mg plant tissue. If used according to the GET™ DNA Template is a spin column format kit suitable for the protocols this kit purifies DNA from 20gm plant tissue. preparation of DNA templates from blood, cells, fungal and animal The kit uses a rapid precipitation technique that uses unique tissue samples. The method involves the solubilization of samples in precipitation reagents to isolate genomic DNA free from proteins and the supplied Template Extraction Buffer, following removal of cellular RNA. Pure genomic DNA is isolated in ~20-40 minutes. debris and proteins the DNA is selectively bound to a high efficiency FEATURES GET™ Spin Column. The pure, genomic DNA is washed and eluted • High Yield, ~100kb genomic DNA from the column in a small volume of an elution buffer. The isolated genomic DNA is suitable for PCR and other applications. • A260/A280 between 1.8 and 2.0 • Simple two tube method, 20-40 minutes • Overcomes the high concentration of plant polysaccharides • No toxic chemicals, no phenol, no hazardous waste APPLICATIONS • Extraction of pure genomic DNA from plant tissues • Suitable for fresh or frozen plant tissues

Cat. No. Description Size 95029-208 OmniPrep™ for Plant For 20gm plant tissue Figure 2: General scheme for GET™ DNA Template and GET™ Plant DNA. GET™ DNA Template purifies mammalian genomic DNA free from OmniPrep™ for Mouse Tail impurities known to inhibit downstream applications. The high yield, ———————————————————————————— extracted DNA is normally >50kb in size. The yield is ~1-40µg DNA High quality genomic DNA from mouse tail per preparation, depending on the source and quantity used. The kit is supplied with reagents for extracting DNA from 50 or 100 preps. OmniPrep™ for Mouse Tail kit isolates high quality genomic DNA FEATURES from mouse tail samples. The kit isolates high purity (A260/A280 ratios of 1.8 to 2.0) DNA (~100kbp) and the yield is 70-80μg/cm tail. If • Spin column format used according to the protocols this kit purifies DNA from 100-200cm • For extraction of genomic DNA from blood, cells fungal and animal mouse tail. tissue The kit uses a rapid precipitation technique that uses unique • Compatible with PCR and other downstream applications precipitation reagents to isolate genomic DNA free from proteins Cat. No. Description Size ™ and RNA. This kit is supplied with additional LongLife Proteinase K 95057-564 GET™ DNA Template 50 preps for the rapid release of DNA from the tough mouse tail tissue. Pure 95057-566 GET™ DNA Template 100 preps genomic DNA is isolated in 20-40 minutes. FEATURES ™ • High Yield, ~100kb genomic DNA GET Plant DNA Template • A /A between 1.8 and 2.0 ———————————————————————————— 260 280 Spin column genomic DNA isolation from plants • Simple two tube method, 20-40 minutes • Supplied with LongLife™ Proteinase K for efficient DNA release A spin column format kit suitable for the preparation of • No toxic chemicals, no phenol, no hazardous waste DNA templates from plant samples. The method involves the solubilization of plant tissue in the supplied Template Extraction APPLICATIONS Buffer, following removal of cellular debris and proteins the DNA is • Extraction of pure genomic DNA from mouse tail tissue selectively bound to a high efficiency GET™ Spin Column. The pure, • Extraction of high quality, 100kb genomic DNA genomic DNA is washed and eluted from the column in a small Cat. No. Description Size volume of an elution buffer. The isolated genomic DNA is suitable for PCR and other applications. ™ 95029-216 OmniPrep for Mouse Tail For 200cm tail Purifies plant genomic DNA free from plant impurities known to inhibit downstream applications. The high yield, extracted DNA is normally >50kb in size. The yield is ~30µg DNA per preparation, depending on the plant source and quantity used. The kit is supplied with reagents for extracting DNA from 100 x 50-100mg plant tissue. FEATURES • Spin column format • For extraction of genomic DNA from plant tissue • Compatible with PCR and other downstream applications

Cat. No. Description Size 95057-568 GET™ Plant DNA Template 100 preps

4 For further details, visit VWR.com Genomic DNA Purification Protein Free DNA with XIT™ XIT™ DNA from Cells ———————————————————————————— ———————————————————————————— Genomic DNA isolation kits that allow you to produce protein free, The XIT™ DNA from Cells kit is designed for the isolation of high quality DNA through the principle of cell lysis, protein digestion genomic DNA from cultured cells. and precipitation, and finally DNA precipitation to isolate high quality The kit uses rapid & efficient nuclear lysis and complete genomic DNA. High quality DNA can be isolated from sample types protein precipitation, followed by DNA precipitation. The yield is including: approximately 5-10µg/1-2x106 cells. · Animal tissues · Cells · Whole blood FEATURES · Bacteria · Buccal cells · Plant tissues • Isolate high quality genomic DNA · Mouse Tail · Yeast · FFPE tissue • Suitable for adherent or suspension cultured cells The XIT™ kits procedure removes contaminants and enzyme • Yield 5-10µg/1-2x106 cells inhibitors, allowing the purified DNA to be ready for immediate use for • Suitable for archive quality DNA all downstream analyses. Cat. No. Description Size ™ XIT DNA from Tissue 95057-638 XIT™ DNA from Cells For 5x107 cells ———————————————————————————— 95057-640 XIT™ DNA from Cells For 5x108 cells The XIT™ DNA from Tissue kit is designed for the isolation of genomic DNA from fresh, frozen or methanol/ acetone fixed tissues. ™ The kit uses three main steps that are rapid & efficient cell lysis, XIT DNA from Blood enzymatic protein digestion and complete protein precipitation, ———————————————————————————— Designed for the isolation of genomic DNA from blood, bone followed by DNA precipitation. This yields high quality genomic DNA. marrow and buffy coat. The kit is supplied with three protocols for the isolation of: The kit uses four main steps that are red blood cell lysis, rapid • 1-10mg and 50-100mg fresh or frozen tissue & efficient nuclear lysis, enzymatic protein digestion and complete • 5-10mg fixed tissue protein precipitation, followed by DNA precipitation. This yields high Three sizes of kit are available for processing a total of 0.25, 2.5 quality genomic DNA. and 25g of tissue. The purified DNA has a A /A ratio between 260 280 Three sizes of kit are available for processing a total of 12.5, 125 1.7 and 1.9 and is up to 200kb in size. Yield is approximately 0.5- or 250ml of blood. The purified DNA has a A /A ratio between 10µg/mg tissue. 260 280 1.7 and 1.9 and is up to 200kb in size. The yield is approximately 10-15µg/ml of blood.

Figure 3: XIT™ DNA from Tissue scheme. FEATURES • Isolate high quality genomic DNA ™ • Suitable for fresh, frozen and fixed tissue Figure 5: XIT DNA from Blood scheme. • Yield 0.5-10µg/mg tissue FEATURES • Suitable for archive quality DNA • Isolate high quality genomic DNA • Suitable for blood, bone marrow and buffy coat • Yield 10-15µg/ml blood • Suitable for archive quality DNA

Figure 4: 10mg mouse liver was treated with the XIT™ Genomic DNA from Tissue kit. The precipitated DNA was hydrated in 30µl TE buffer and 3µl was loaded onto an agarose gel. Figure 6: Genomic DNA was isolated from 500µl bloodusing the XIT™ Cat. No. Description Size Genomic DNA from Blood Kit. 95057-658 XIT™ DNA from Tissue For 250mg tissue 95057-660 XIT™ DNA from Tissue For 2.5g tissue Cat. No. Description Size 95057-662 XIT™ DNA from Tissue For 10g tissue 95057-628 XIT™ DNA from Blood For 12.5ml blood 95057-630 XIT™ DNA from Blood For 125ml blood 95057-632 XIT™ DNA from Blood For 250ml blood

For further details, visit VWR.com 5 Genomic DNA Purification XIT™ DNA from Buccal Cells XIT™ DNA from Mouse Tail ———————————————————————————— ———————————————————————————— Designed for the isolation of genomic DNA from Buccal cells The XIT™ DNA from Mouse Tail kit is designed for the isolation of (cheek cells). The kit uses three main steps that are rapid & genomic DNA from 5mm sections of mouse tail. The kit uses three efficient cell lysis, enzymatic protein digestion and complete protein main steps rapid & efficient cell lysis with enzymatic protein digestion precipitation, followed by DNA precipitation. This yields high quality and complete protein precipitation, followed by DNA precipitation. genomic DNA. This yields high quality DNA. The kit is available with cheek cell collection brushes or a The kit is available for processing a total of 125mm of mouse tail.

mouthwash method to collect Buccal cells. The purified DNA has a 260A /A280 ratio between 1.7 and 1.9 and is up The kits are available in two different sizes. The purified DNA has to 200kb in size.

a A260/A280 ratio between 1.8 and 2.0 and is up to 200kb in size. The yield is approximately 3-5µg/swab.

Figure 10: XIT™ DNA from Mouse Tail scheme. Figure 7: XIT™ DNA from Buccal Cells scheme. FEATURES Figure 8: Cheek cells were collected with the supplied • Isolate high quality genomic DNA collection brush and the genomic DNA extracted with the • Suitable for archive quality DNA XIT™ Genomic DNA from Buccal Cell kit. The isolated genomic DNA was used as a template for PCR amplification. The Cat. No. Description Size resulting PCR products are shown. 95057-668 XIT™ DNA from Mouse Tail For 125mm mouse tail

Cat. No. Description Size ™ 95057-650 XIT™ DNA from Buccal Cells (Mouthwash) For 25 samples XIT DNA from Plant Tissues 95057-652 XIT™ DNA from Buccal Cells (Mouthwash) For 250 samples ———————————————————————————— Designed for the isolation of genomic DNA from fresh or frozen 95057-654 XIT™ DNA from Buccal Cells (Collection Brush) For 25 samples plant tissues. The kit uses three main steps that are rapid & 95057-656 XIT™ DNA from Buccal Cells (Collection Brush) For 50 samples efficient cell lysis, enzymatic protein digestion and complete protein precipitation, followed by DNA precipitation. This yields high quality ™ XIT DNA from FFPE Tissue genomic DNA. ———————————————————————————— Two sizes of kit are available for processing a total of 2.5 and 25g Designed for the isolation of genomic DNA from formalin fixed, of tissue. The purified DNA has a 260A /A280 ratio between 1.7 and paraffin embedded tissue. The kit uses three main steps that are 1.9 and is up to 200kb in size. The yield is around 1-5µg/mg tissue, rapid & efficient cell lysis, enzymatic protein digestion and complete dependent on plant species. protein precipitation, followed by DNA precipitation. This yields high quality genomic DNA. The kit is for processing a total of 0.25g of tissue. The purified

DNA has a A260/A280 ratio between 1.7 and 1.9 and is up to 200kb in size. The yield is approximately 0.5-10µg/mg tissue.

Figure 11: XIT™ DNA from Plant scheme.

Figure 12: Genomic DNA was extracted from 100mg Arabidopsis thaliana with the XIT™ Genomic DNA from Plant Tissue kit. The DNA was rehydrated in 50µl & 10µl was loaded onto an agarose gel. FEATURES Figure 9: XIT™ DNA from FFPE Tissue scheme. • Yield 1-5µg/mg tissue • Suitable for archive quality DNA FEATURES • Suitable for fresh, frozen and fixed tissue • Yield 0.5-10µg/mg tissue • Suitable for archive quality DNA Cat. No. Description Size 95057-634 XIT™ DNA from Plant Tissue For 2.5g tissue Cat. No. Description Size 95057-636 XIT™ DNA from Plant Tissue For 25g tissue 95057-626 XIT™ DNA from FFPE Tissue For 250mg tissue

6 For further details, visit VWR.com Genomic DNA Purification XIT™ DNA from Gram Positive Bacteria XIT™ DNA from Yeast ———————————————————————————— ———————————————————————————— Designed for the isolation of genomic DNA from overnight Designed for the isolation of genomic DNA from overnight yeast cultures. The kit uses three main steps that are rapid & efficient cultures. enzymatic digestion of cell walls, spheroplast lysis and complete The kit uses three main steps that are rapid & efficient enzymatic protein precipitation, followed by DNA precipitation. This yields high digestion of cell walls, spheroplast lysis and complete protein quality genomic DNA. precipitation, followed by DNA precipitation. This yields high quality Two sizes of the kit are available for processing a total of 25 genomic DNA. and 250ml of bacterial culture. The purified DNA has a 260A /A280 Two sizes of kit are available for processing a total of 25 and ratio between 1.7 and 1.9 and is up to 200kb in size. The yield is 250ml of yeast culture. XIT™ DNA from Yeast Kit protocol is designed approximately 15-25µg/ml culture. to use 1ml overnight culture, however the protocol can be easily

adapted for larger tissue sample sizes.The purified DNA has a 260A /

A280 ratio between 1.7 and 1.9 and is up to 200kb in size. The yield is approximately 1-6µg/ml culture.

Figure 13: XIT™ DNA from Gram Positive Bacteria scheme. FEATURES • Yield 15-25µg/ml bacterial culture Figure 15: XIT™ DNA from Yeast scheme. • Suitable for archive quality DNA

Cat. No. Description Size 95057-646 XIT™ DNA from Gram positive bacteria For 25ml culture 95057-648 XIT™ DNA from Gram positive bacteria For 250ml culture

™ XIT DNA from Gram Negative Bacteria Figure 16: Genomic DNA was isolated from 1ml overnight yeast culture, using ———————————————————————————— XIT™ DNA from Yeast kit. The DNA was resuspended in 50µl and 5µl or 10µl Designed for the isolation of genomic DNA from overnight were resolved on an agarose gel. cultures. The kit uses three main steps that are rapid & efficient FEATURES bacterial lysis, complete protein precipitation, followed by DNA precipitation. This yields high quality genomic DNA. • Yield 1-6µg/mg tissue Two sizes of kit are available for processing a total of 25 and • Suitable for archive quality DNA 250ml of bacterial culture. The protocol is designed to use 1ml Cat. No. Description Size overnight culture, however the protocol can be easily adapted for 95057-664 XIT™ DNA from Yeast For 25ml culture larger culture sizes. The purified DNA has a A /A ratio between 260 280 95057-666 XIT™ DNA from Yeast For 250ml culture 1.7 and 1.9 and is up to 200kb in size. The yield is approximately 20-40µg/ml culture. XIT™ Mitochondrial DNA ———————————————————————————— The XIT™ Mitochondrial DNA kit combines our FOCUS™ Mitochondria technology with our XIT™ DNA isolation technology in a single kit. The kit uses our proprietary Subcell buffers to lyse cells and animal tissue and remove cellular proteins and nuceli from a highly enriched, intact and active mitochondrial fraction. The mitochondria are then lyzed with the included lysis buffer. The lysate is treated with our LongLife™ Proteinase K to degrade Figure 14: XIT™ DNA from Gram Negative Bacteria scheme. the proteins, which are then precipitated and the mitochondrial DNA is isolated with alcohol precipitation. FEATURES The kits is designed for ~100 preps consisting of 20x106 • Yield 20-40µg/mg tissue mammalian cells/ prep. The kit protocol also contains protocols for • Suitable for archive quality DNA isolation of mitochondra from soft (i.e. brain & liver) and hard (i.e. Cat. No. Description Size cardiac & skeletal muscle) tissue. 95057-642 XIT™ DNA from Gram negative bacteria For 25ml cu ture FEATURES 95057-644 XIT™ DNA from Gram negative bacteria For 250ml culture • Isolate high quality mitochondrial DNA • Suitable for cultured cells, soft and hard animal tissue

Cat. No. Description Size 89167-750 XIT™ Mitochondrial DNA 100 preps For further details, visit VWR.com 7 Genomic DNA Purification PCR Template DNA Rapid DNA Template Prep™ ———————————————————————————— ———————————————————————————— For small sample size with low DNA content ™ OmniTemplate This kit is suitable for the preparation of DNA templates from ———————————————————————————— blood, cells, animal tissues and plant samples. The method involves Single tube preparation of PCR ready genomic DNA solubilization of a sample in Template Extraction Buffer, followed by templates the selective binding of DNA to our proprietary pinkRESIN™. Following ™ OmniTemplate™ is specifically designed for the rapid isolation of a washing step, the DNA template is eluted from the pinkRESIN . The a DNA template for polymerase chain reaction (PCR) analysis from isolated template is suitable for PCR and other applications. One mammalian tissue samples, blood and cell cultures. This single tube preparation is for 1-10mg animal tissue or 50-100mg plant tissue. method produces a high concentration, ready-to-use supply of DNA template for microanalysis, genotyping and a multitude of other applications. The method involves isolation of nuclei and their subsequent digestion with LongLife™ Proteinase K in the same tube the samples were collected in. The isolated nuclei are digested to release template DNA in a buffer compatible with PCR and other downstream applications. The OmniTemplate™ samples can be directly used in PCR and other applications without further manipulations. One prep is approximately 1-10mg tissue.

Figure 18: Rapid DNA Template Prep™ scheme. FEATURES • For animal and plant samples • Compatible with PCR and other downstream applications APPLICATIONS • For the rapid preparation of DNA template

Cat. No. Description Size 82022-368 Rapid DNA Template Prep™ 50 preps Figure 17: OmniTemplate™ produces high quality genomic DNA template. 82022-370 Rapid DNA Template Prep™ 100 preps 5μl (lanes 1, 3, 5) or 15µl (lanes 2, 4, 6) PCR reactions were loaded onto a 2% agarose gel. Template was purified from 104 (lanes 1-2), 106 (lanes 3-4) or 2.8x106 (lanes 5-6) NIH3T3 cells. Yeast Geno-DNA-Template™ ———————————————————————————— FEATURES Extraction of high quality DNA from yeast • Single tube genomic DNA isolation • Compatible with PCR and other downstream applications This extraction kit isolates high quality genomic DNA from yeast. ™ • Suitable for blood, cells, and animal tissue samples The kit is based on a two-step lysis of yeast cells, using LongLife ® ™ • No toxic chemicals, no phenol, no hazardous waste Zymolyase and LongLife Proteinase K, followed by the removal of proteins and other cellular impurities by precipitation and APPLICATIONS centrifugation. The clean, genomic DNA is collected by precipitation • Designed for the rapid preparation of DNA template for large and the high yield of extracted DNA has an average 100kb length and throughput microanalysis has A260/280 1.8-2.0. • High recovery of ready-to-use DNA templates in a short period of For extracting DNA from 100 x 1.5ml yeast cultures. time

Cat. No. Description Size 82023-256 OmniTemplate™ >100 preps

Figure 19: Yeast Geno-DNA-Template™ scheme. FEATURES • Compatible with PCR and other downstream applications APPLICATIONS • For the preparation of high yield, DNA template from yeast

Cat. No. Description Size 82021-534 Yeast Geno-DNA-Template™ 100 preps 8 For further details, visit VWR.com Genomic DNA Purification Plant Geno-DNA-Template™ High Molecular Weight DNA ———————————————————————————— ———————————————————————————— Extraction of high quality DNA from plants ™ Plant-Geno-DNA-Template™ is suitable for the preparation of DNA MegaLong templates from plant samples. The method involves the solubilization ———————————————————————————— of plant tissue in the supplied Template Extraction Buffer, following Extract high molecular weight genomic DNA with removal of cellular debris and proteins the DNA is selectively bound minimal shearing to pinkRESIN™. The pure, genomic DNA is washed and eluted from ™ The majority of genomic DNA extraction methods involve the pinkRESIN in a small volume of an elution buffer. The isolated numerous physical manipulations, including mixing, pipetting, genomic DNA is suitable for PCR and other applications. ™ shaking, binding to resin and elution, which results in sheared DNA Plant Geno-DNA-Template purifies plant genomic DNA free from that may not be suitable for further analysis. plant impurities known to inhibit downstream applications. The high MegaLong™ isolates high molecular weight genomic DNA from a yield, extracted DNA is normally >50kb in size. variety of samples, including animal tissues, cultured cells, whole The kit is supplied with reagents for extracting DNA from 100 x blood, bacterial and yeast. MegaLong™ uses Genomic Tube-O- 50-100mg plant tissue. DIALYZER™, a unique, patented micro dialysis device with a 0.45μm membrane that minimizes sample manipulation, one of the main reasons for DNA breakage. Nuclei are isolated under mild extraction conditions and genomic DNA is released by digestion of nuclear proteins with a highly active LongLife™ Proteinase K. The digestion is performed in the Tube-O-DIALYZER™ and after digestion the Tube- O-DIALYZER™ is inverted to dialyze away digested protein and other impurities leaving behind highly pure and fully hydrated genomic DNA. The fragile, high molecular weight genomic DNA can be stored in the Tube-O-DIALYZER™ to further minimize mechanical manipulation of the DNA. The DNA is suitable for Southern blot analysis, recovery of Lambda shuttle vectors from transgenic animals, PCR, analysis by pulsed-field electrophoresis or any application where genomic DNA is ™ Figure 20: Plant Geno-DNA-Template scheme. required. FEATURES The kit is supplied in two sizes for the purification of either 25 or 50 x 1-100mg samples. • For extraction of genomic DNA from plant tissue • Compatible with PCR and other downstream applications APPLICATIONS • Designed for the preparation of high yield, genomic DNA template from plant

Cat. No. Description Size 82021-536 Plant Geno-DNA-Template™ 100 preps

Figure 21: Megalong™ scheme. FEATURES • Isolation of high molecular weight genomic DNA • For animal tissues, cultured cells, whole blood, bacterial and yeast • Uses patented genomic Tube-O-DIALYZER™ to minimize sample manipulation APPLICATIONS • For DNA libraries, PCR amplification, Southern blot analysis, pulse- field electrophoresis, and other applications where >100kb DNA is required

Cat. No. Description Size 82021-542 MegaLong™ 25 preps 82021-544 MegaLong™ 50 preps

For further details, visit VWR.com 9 Plasmid DNA Tools Plasmid Isolation GET™ Plasmid DNA Mediprep ———————————————————————————— ———————————————————————————— Spin column plasmid isolation from 25-200ml GET™ Plasmid DNA Miniprep culture ———————————————————————————— Spin column plasmid isolation from 1-5ml culture; Isolates high quality plasmid DNA from 25-200ml E. coli cultures. The kit utilizes an enhanced DNA binding column to produce high single column or 96-well. yields of plasmid. This quick and easy protocol eliminates toxic Isolates high quality plasmid DNA from 1-5ml E. coli cultures. phenol/chloroform extractions or lengthy ethanol precipitations. On The kit utilizes an enhanced DNA binding column to produce high completion of the protocol, the plasmid DNA is ready for restriction yields of plasmid. This quick and easy protocol eliminates toxic enzyme digestion, sequencing, subcloning and in vitro transcription. phenol/chloroform extractions or lengthy ethanol precipitations. On Plasmid yields are typically up to 20-100µg/prep. completion of the protocol, the plasmid DNA is ready for restriction FEATURES enzyme digestion, sequencing, subcloning and in vitro transcription. • Isolate up to 100μg ready-to-use plasmid DNA Plasmid yields are typically up to 20µg/prep. Available as single use • No phenol, chloroform or alcohol precipitation spin columns or a 96-well spin format. • 25 and 50 prep kits available.

Cat. No. Description Size 95057-578 GET™ Plasmid Mediprep 25 preps 95057-580 GET™ Plasmid Mediprep 50 preps GET™ Plasmid DNA Maxiprep ———————————————————————————— Spin column plasmid isolation from 100-500ml culture Isolates high quality plasmid DNA from 100-500ml E. coli cultures. The kit utilizes an enhanced DNA binding column to produce high yields of plasmid. This quick and easy protocol eliminates toxic phenol/chloroform extractions or lengthy ethanol precipitations. On completion of the protocol, the plasmid DNA is ready for restriction enzyme digestion, sequencing, subcloning and in vitro transcription. Plasmid yields are typically up to 100-500µg/prep. FEATURES • Isolate up to 500μg ready-to-use plasmid DNA • No phenol, chloroform or alcohol precipitation • 10 and 20 prep kits available.

Cat. No. Description Size 78000-058 GET™ Plasmid Maxiprep 10 preps 78000-060 GET™ Plasmid Maxiprep 20 preps

Figure 22: GET™ Plasmid DNA single and 96-well plate general scheme.

Figure 23: Comparison of GET™ Plasmid Miniprep with Competitor Q. Overnight cultures were processed with G-Biosciences’ GET™ or Competitor Q’s Plasmid Miniprep kit. DNA was eluted in an equal volume of TE buffer and resolved. Both kits produced a strong band of supercoiled plasmid DNA and some minor higher bands due to nicked DNA, however Competitor Q had additional bands below the main band that are denatured plasmid DNA that is resistant to restriction digestion. FEATURES • Isolate up to 20μg ready-to-use plasmid DNA • Spin or vacuum manifold compatible • No phenol, chloroform or alcohol precipitation • 50 and 100 prep kits available.

Cat. No. Description Size 78000-054 GET™ Plasmid Miniprep 50 preps 78000-056 GET™ Plasmid Miniprep 100 preps 89130-996 GET™ Plasmid DNA 96-well 4 x 96-well plates 10 For further details, visit VWR.com Plasmid DNA Tools Bacteria Transformation Plasmid Screening ———————————————————————————— ———————————————————————————— Z-Competent™ E. coli Transformation Plasmid Screening ToothPick™ ———————————————————————————— ———————————————————————————— High efficiency competent cells in <20 minutes Rapid colony screening for plasmid DNA Designed to generate competent E. coli cells for simple and highly Plasmid Screening ToothPick™ is a unique system that allows for efficient E. coli transformation. the rapid screening of bacteria for transformed plasmids, without the E. coli are grown in SOB medium, washed and suspended need for an overnight culture. in the supplied Competent Buffer. The bacterial cells are now Pick a colony, add to Plasmid Screening ToothPick™ reagents and ready for transformations. The transformation efficiency is 2x108- then analyze using restriction enzymes. 1x109 transformants per μg of pUC19 plasmid. The efficiency of transformation varies on E. coli strain and plasmid size. The competent cells can be used immediately or stored for later use. The kit is supplied with reagents sufficient for processing up to 300 x 100μl aliquots of competent cells for transformations or with reagents and SOB culture media for processing up to 100 x 100μl aliquots of competent cells.

FEATURES Figure 24: ToothPick™ general scheme. • No heat shock required • Competent cells in <20mins FEATURES • Transformations in <5mins • Simple, 5 minute protocol, as easy as picking a colony APPLICATIONS • No overnight cultures required • Generation of competent cells from E. coli APPLICATIONS • Rapid transformation of competent cells • Rapid colony screening, using restriction digestion analysis CITED REFERENCES Cat. No. Description Size Singh, S. et al (2005) Protein Sci. 14: 2095 82022-372 Plasmid Screening ToothPick™ 300 preps Singh, S. et al (2005) Protein Sci. 14: 2601 Tyler, R. et al (2005) Proteins. 59: 633 ™ Cat. No. Description Size ToothPick -PCR ™ ———————————————————————————— 82023-074 Z-Competent E.coli Transformation 300 preps Rapid colony screening for plasmid DNA with PCR Z-Competent™ E.coli Transformation 82023-076 100 preps ™ ™ with SOB media ToothPick -PCR is an extension of ToothPick and allows for the 82023-078 SOB Media 50ml rapid release of plasmids from transformed bacteria for screening by polymerase chain reaction (PCR). There is no requirement for growing bacteria, performing “minipreps” or purifying the plasmid DNA. Plasmid Screening Reagents ™ ———————————————————————————— Add your colony directly to the supplied ToothPick solution, heat Antibiotics, IPTG, X-Gal and X-Gluc and then transfer 1µl directly to a PCR reaction to screen for your plasmid. Following PCR, add the supplied GLOW™ Loading Dye and HP Ampicillin™ is a stronger and more stable inhibitor of run on an agarose gel. No need to stain with ethidium bromide, β-lactamase than regular ampicillin and, therefore, supports only simply place gel on UV box and view. the growth of ampicillin resistant colonies. HP Ampicillin™ is supplied Glow Dyes™ have been specifically formulated for nucleic acid as ten lyophilized vials suitable for 500ml/vial. Regular ampicillin is electrophoresis and contain an optimized concentration of buffer also offered. agents, tracking dyes and ethidium bromide. These loading dyes IPTG induces activity of β-galactosidase by binding and inhibiting reduce exposure and many of the problems associated with ethidium the lac repressor. In cloning experiments, the lacZ gene is replaced bromide use and disposal. with the gene of interest and IPTG is then used to induce gene expression. IPTG is an effective inducer in the concentration range of 100μM to 1.5mM X-gal is used to indicate whether bacteria express the β-galactosidase enzyme, which is encoded by the lacZ gene. X-Gluc is a substrate for β-glucuronidase (GUS), which is encoded by gusA, a widely used reporter gene. CITED REFERENCES Figure 25: Screening of Plasmids with ToothPick™-PCR. Four transformed Oatley, M. et al (2011) Biol Reprod. 85:347 bacterial colonies were picked, resuspended in 15µl ToothPick™ solution & boiled for 5 minutes. 1µl was added to a 50µl standard PCR reaction & the Cat. No. Description Size resulting PCR products were resolved on an agarose gel. Lanes 1 & 2 show ™ 95029-318 HP Ampicillin [1000X] For 5L the correct plasmid. 71003-352 Ampicillin sodium salt 25g FEATURES 71003-354 Ampicillin sodium salt 100g • Simple, 5 minute protocol, as easy as picking a colony 82023-106 IPTG (Molecular Biology Grade) 5g • No overnight cultures required 82023-104 X-Gal (Molecular Biology Grade) 1g • Supplied with GLOW™ Loading Dye 95029-328 X-Gluc (Molecular Biology Grade) 100mg Cat. No. Description Size 95029-222 ToothPick™-PCR 300 preps For further details, visit VWR.com 11 DNA Cleanup & Concentration GET™ CLEAN DNA Genomic Tube-O-DIALYZER™ ———————————————————————————— ———————————————————————————— For PCR clean up & removal of restriction enzymes. For the rapid clean-up of genomic DNA GET™ CLEAN DNA uses our high binding affinity GET™ Spin Genomic Tube-O-DIALYZER™ rapidly dialyzes >100kb genomic DNA Columns to remove salts, enzymes, unincorporated nucleotides, samples to remove small DNA, salts, RNA and other contaminants radiolabels, and primer-dimers from any DNA preparation of 100bp to with minimal hands-on manipulation. >20kb. GET™ Spin Columns has an enhanced binding affinity for DNA, Based on our patented Tube-O-DIALYZER™, the genomic Tube- thus eliminating loss or damage of DNA. O-DIALYZER™ has a dialysis membrane secured in the cap. Simply The DNA cleaning protocol takes as little as 5 minutes: pipette your >100kb genomic DNA sample into the Tube-O-DIALYZER™ GET™ CLEAN DNA is available in a 50 or 100 prep size. tube, screw on the dialysis cap and dialyze. To recover 100% of your sample, briefly centrifuge and replace the dialysis cap with the supplied storage cap and store.

Figure 26: GET™ CLEAN DNA scheme. FEATURES • Spin column format • Enhanced DNA binding affinity Figure 27: Genomic Tube-O-DIALYZER™ scheme. • Protocol is 5-10 minutes • No phenol/chloroform extraction or alcohol precipitations FEATURES APPLICATIONS • Rapid removal of impurities, RNA, small fragment DNA, etc. • PCR clean-up • 100% sample recovery • Purifying DNA fragments after in-vitro labeling reactions • Single tube for dialysis and storage, prevents sample loss • Removal of restriction enzymes from plasmid DNA • For 0.2-2.5ml sample • Cleaning any DNA preparations from interfering agents • No contamination APPLICATIONS Cat. No. Description Size • For the clean-up of genomic (>100kb) DNA by dialysis ™ 95057-570 GET CLEAN DNA 25 preps • Patented, single tube dialysis device to ensure no sample loss 95057-572 GET™ CLEAN DNA 50 preps Cat. No. Description Size pinkCLEANUP™ 82022-546 Genomic Tube-O-DIALYZER™ 25 ———————————————————————————— For the rapid clean up of DNA preparations Spin-OUT™ for PCR Uses pinkRESIN™ to remove salts, enzymes, unincorporated ———————————————————————————— nucleotides, radiolabels, and primer-dimers from any DNA Remove contaminants after PCR preparations of 100bp to >20kb. pinkRESIN™ has an enhanced The SpinOUT™ PCR columns are spin format desalting columns binding affinity for DNA and resuspends with ease, thus eliminating that have the ability to remove salts, radioisotpes, dyes, primers and loss or damage of DNA; a major frustration of using binding matrix for deoxynucleotides (dNTPs) for nucleic acids following PCR. Two sizes isolation of DNA. are available to remove contaminating products from PCR products, The DNA cleaning protocol takes as little as 5 minutes. including <20bp primers, dNTPs and salts or from <32bp primers, Optional spin columns are also offered to simplify the procedure dNTPs and salts. further. PinkRESIN™ is also offered separately for those who prefer to Spin-OUT™ PCR is for the cleaning of PCR products. PCR-20 customize their own protocol. removes contaminating products from PCR products, including FEATURES <20bp primers, dNTPs and salts. PCR-32 removes PCR products from <32bp primers, dNTPs and salts. • Enhanced DNA binding affinity and rapidly resuspends • Protocol is 5-10 minutes FEATURES • No phenol/chloroform extraction or alcohol precipitations • Available for <20bp or <32bp primers APPLICATIONS • Spin format for rapid purification • PCR clean-up, purifying after in-vitro labeling, restriction enzyme APPLICATIONS removal or cleaning any DNA preps • Remove primers, dNTPs and other decontaminants from PCR reactions Cat. No. Description Size 82022-734 pinkCLEANUP™ 200 preps Cat. No. Description Size 82022-736 pinkRESIN™ 1ml 82022-588 Spin-OUT™ PCR-20 10 columns 82022-590 Spin-OUT™ PCR-32 10 columns

12 For further details, visit VWR.com DNA Cleanup & Concentration Spin-OUT™ GET™ AGAROSE DNA ———————————————————————————— ———————————————————————————— For desalting and buffer exchange Rapid purification of DNA from agarose gels The SpinOUT™ GT-600 and GT-1200 columns are versatile, spin- This kit is based on our GET™ Spin Columns, spin columns with a format columns for the desalting and buffer exchange of nucleic high binding affinity for DNA. acid solutions ranging from 5µl through to 4ml sample volumes. The GET™ AGAROSE DNA method involves release of DNA fragments SpinOUT™ columns are available in two MWCO sizes for nucleic acids from gel pieces followed by capture of nucleic acids on the GET™ Spin larger than 10bp or nucleic acids larger than 20bp. Columns, washing, and elution of the clean nucleic acid fragments in The SpinOUT™ columns are simply to use as the sample is applied a suitable buffer. and then centrifuged to recover nucleic acids with the column The GET™ AGAROSE DNA kits are supplied with enough reagents retaining >95% of the salts and small molecules (<1,000Da). for 50 or 100 preps. Spin-OUT™ GT-600 is for the purification of nucleic acids larger than 10bp and proteins > 6kDa. Spin-OUT™ GT-1200 is for the purification of proteins >30bp and the removal of molecules >1,500Da. Five sizes are available for each MWCO and are highlighted below: FEATURES • 5 sizes available for sample volumes of 5µl to 4ml • Spin format for rapid purification Figure 28: GET™ AGAROSE DNA scheme. CITED REFERENCES Tripodi, K et al (2005) Plant Physiol. 139:969 FEATURES Taggert, C. et al (2005) J. Exp. Med. 202:1659 • Extract DNA fragments from agarose gels Resin • Rapid DNA isolation Bed Sample • DNA ready for downstream applications, including ligations Cat. No. Description Size (ml) Load (ml) • Suitable for 100-20,000bp fragments 89167-920 SpinOUT™ GT-600, 0.1ml 25 columns 0.1 0.005-0.02 • Compatible with TAE and TBE buffer gels 82022-580 SpinOUT™ GT-600, 1ml 10 columns 1 0.05-0.1 APPLICATIONS 82022-582 SpinOUT™ GT-600, 3ml 10 columns 3 0.1-0.5 • Isolation of DNA fragments from agarose gels 89167-922 SpinOUT™ GT-600, 5ml 5 columns 5 0.5-2 • Cleaning and removing of contaminants from DNA samples ™ 89167-924 SpinOUT GT-600, 10ml 5 columns 10 0.5-4 Cat. No. Description Size ™ 89167-926 SpinOUT GT-1200, 0.1ml 25 columns 0.1 0.005-0.02 95057-574 GET™ AGAROSE DNA 50 preps ™ 82022-584 SpinOUT GT-1200, 1ml 10 columns 1 0.05-0.1 95057-576 GET™ AGAROSE DNA 100 preps 82022-586 SpinOUT™ GT-1200, 3ml 10 columns 3 0.1-0.5 89167-928 SpinOUT™ GT-1200, 5ml 5 columns 5 0.5-2 ™ ™ geneEXIT 89167-930 SpinOUT GT-1200, 10ml 5 columns 10 0.5-4 ———————————————————————————— Isolation of nucleic acids from agarose gels The kit is based on pinkRESIN™, a high capacity, proprietary binding resin matrix for nucleic acids. pinkRESIN™ has an enhanced binding affinity for DNA and RNA and resuspends with ease, thus eliminating loss and damage of nucleic acids, a frequent problem with other binding resin matrices for DNA and RNA isolation. pinkRESIN™ is a non-toxic matrix. geneEXIT™ method involves release of nucleic acid fragments from gel pieces followed by capture of nucleic acids with pinkRESIN™, washing, and elution of the clean nucleic acid fragments in a suitable buffer. The geneEXIT™ method can be carried out with or without the use of spin columns; however the use of spin columns simplifies the protocols. FEATURES • Rapid nucleic acid isolation • DNA & RNA ready for further applications, including ligations • Suitable for 100-20,000bp fragments • Compatible with TAE and TBE buffer gels • Supplied with or without spin columns APPLICATIONS • Isolation of DNA/RNA fragments from agarose gels • Cleaning contaminants from DNA & RNA samples

Cat. No. Description Size 82022-730 geneEXIT™ 100 preps 82022-732 geneEXIT™ with spin columns 100 preps

For further details, visit VWR.com 13 PCR/ RT-PCR DNA Polymerases Pfu Polymerase ———————————————————————————— ———————————————————————————— Pfu DNA polymerase, derived from the hyperthermophilic archae Taq Polymerase Pyrococcus furiosus, has superior thermostability and proofreading ———————————————————————————— properties compared to other thermostable polymerase. Its molecular A recombinant Taq DNA Polymerase is a highly thermostable weight is 90 kDa. It can amplify DNA target up to 2kb (simple DNA polymerase isolated from the thermophile, Thermus aquaticus. template). The elongation velocity is 0.2~0.4kb/min(70~75°C). Pfu Taq DNA polymerase catalyzes the 5’>3’ synthesis of DNA. The DNA polymerase possesses 3' to 5' exonuclease proofreading activity enzyme has no detectable 3’>5’ proofreading exonuclease activity, that enables the polymerase to correct nucleotide-misincorporation and possesses low 5’>3’ exonuclease activity. The Taq polymerase errors. This means that Pfu DNA polymerase-generated PCR is able to amplify DNA up to 6kb that have a single adenine 3’ fragments will have fewer errors than Taq-generated PCR inserts. overhang. The error rate of this Taq polymerase is ~2.2x10-5 Using Pfu DNA polymerase in your PCR reactions results in blunt- nucleotide-1 cycle-1. ended PCR products, which are ideal for cloning into blunt-ended Unit Definition vectors. Pfu DNA polymerase is superior for techniques that require high-fidelity DNA synthesis. Supplied with 10X Buffer and 10X One unit is defined as the amount of the enzyme required to Loading Buffer. catalyze the incorporation of 10nmole of dNTPs into an acid-insoluble form in 30 minutes at 70°C using herring sperm DNA as substrate. Unit Definition Storage Buffer One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10nmole of dNTPs into an acid-insoluble 20mM TrisCl ( pH8.0), 100mM KCl, 3mM MgCl , 1mM DTT, 0.1% 2 form in 30 minutes at 70°C using herring sperm DNA as substrate. Nonidet® P-40, 0.1% Tween® 20, 0.2mg/ml BSA, 50% (v/v) glycerol Storage Buffer 10X PCR Buffer 20mM TrisCl (pH8.0), 100mM KCl, 3mM MgCl 1mM DTT, 0.1% 120mM Tris-HCl(pH 8.8), 500mM KCl, 1% Triton® X-100, 100mM 2 Nonidet® P-40, 0.1% Tween® 20, 0.2mg/ml BSA, 50% (v/v) glycerol Lycine 10X Pfu Buffer FEATURES 200mM Tris-HCl(pH8.8@25°C), 100mM KCl, 100mM (NH )2SO , • 5’>3’ polymerase activity 4 4 20mM MgSO , 1.0% Triton® X-100, 1mg/ml BSA • No detectable 3’>5’ proofreading exonuclease activity 4 • Low 5’>3’ exonuclease activity FEATURES • Error rate ~2.2x10-5 nucleotide-1 cycle-1 • 5’>3’ polymerase activity • Available as a 2X Mastermix • 3’>5’ proofreading exonuclease activity • Available as a 2X Mastermix Cat. No. Description Size FEATURES 95057-686 Taq Polymerase 1,000U • High-fidelity PCR and primer-extension reactions Taq Polymerase (2X MasterMix) • High fidelity PCR for cloning into blunt-ended vectors ———————————————————————————— • Site-directed mutagenesis Taq Polymerase is available as a ready-to-use mixture of the Cat. No. Description Size polymerase enzyme, deoxynucleotides (dNTPs) and a 2X reaction 71003-296 Pfu Polymerase 500U buffer. The 2X MasterMix contains all the necessary reagents for successful amplification, except for the DNA template and primers. Pfu Polymerase (2X MasterMix) FEATURES ———————————————————————————— • Ready to use mastermix, just add template & primers Pfu Polymerase (2X MasterMix) is a premixed, ready-to-use

• 5’>3’ polymerase activity solution containing Pfu DNA Polymerase, dNTPs, MgSO4 and • No detectable 3’>5’ proofreading exonuclease activity Reaction Buffer at optimal concentrations for efficient amplification • Error rate ~2.2x10-5 nucleotide-1 cycle-1 of DNA templates by PCR. To prepare the final PCR, only primers and template DNA are added. Pfu Polymerase (2X MasterMix) contributes Cat. No. Description Size to highly reproducible PCR by reducing the risk of pipetting errors, 89167-762 2X MasterMix Taq Polymerase 100 reactions miscalculation and contamination. It also contributes to higher sensitivity by adding intensifier and optimizer. FEATURES • 5’>3’ polymerase activity • 3’>5’ proofreading exonuclease activity • Convenient: Pfu DNA Polymerase in a ready-to-use Mix • High yields of PCR products with minimal optimization • Fast: Saves time due to reduced number of pipetting steps • Reproducible: Lower contamination and pipetting error risk

Cat. No. Description Size 71003-298 Pfu Polymerase (2X MasterMix) 40 reactions

14 For further details, visit VWR.com PCR/ RT-PCR Deoxynucleotides Reverse Transcriptases (RT) ———————————————————————————— ———————————————————————————— Deoxynucleotides (dNTPs) Set M-MLV Reverse Transcriptase ———————————————————————————— ———————————————————————————— The set consists of 100mM aqueous solutions of dATP, dCTP, Moloney Murine Leukemia Virus (M-MLV) Reverse Transcriptase is dGTP and dTTP each supplied in a separate vial. a RNA-dependent DNA polymerase. This enzyme is able to use RNA Since the nucleotides are provided separately, the dNTP Set offers molecule as a template and synthesize single-strand DNA. maximum flexibility in preparation of reaction mixes for different Unit Definition applications. One unit is defined as the amount of enzyme required to Each deoxynucleotide is also available separately at a 100mM incorporate 1nmol of dTTP into acid-precipitable material in 10 min concentration. + at 37°C using poly(A) RNA and oligo(dT)18 as template/primer. ApplicationS Supplied with Enzyme • PCR, long PCR, RT-PCR, cDNA synthesis, primer extension, DNA • 5× Reaction Buffer: 250mM Tris-HCl, 150mM KCl, 40 mM MgCl2, sequencing, DNA labeling pH8.3 Cat. No. Description Size • 100 mM DTT 95057-688 dNTP Set [100mM] 4 x 0.25ml Storage Condition 71003-178 dATP [100mM] 0.25ml 20 mM Tris-HCl, 150 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1% 71003-180 dCTP [100mM] 0.25ml IGEPAL CA-630, 50% Glycerol, pH7.6. Store at -20°C. 71003-182 dGTP [100mM] 0.25ml Application 71003-184 dTTP [100mM] 0.25ml • Synthesis cDNA from a single-stranded RNA or DNA Primer extension Deoxynucleotides (dNTPs) Mix • Sequencing dsDNA ———————————————————————————— • cDNA library Aqueous mixture of dATP, dCTP, dGTP and dTTP. The dNTP Mix is a • Template production for use in RT-PCR ready-to-use aqueous solution containing dATP, dCTP, dGTP and dTTP, • 3'-end labeling of duplex DNA via end-filling reactions each at a final concentration of 10mM. The Mix reduces the number Cat. No. Description Size of pipeting steps and the risk of errors. 71003-284 M-MLV Reverse Transcriptase 10,000U Applications • Ready to use in PCR, long-PCR, RT-PCR, cDNA synthesis, primer AMV Reverse Transcriptase extention and DNA labeling. ———————————————————————————— Cat. No. Description Size Purified from Avian Myeloblastosis Virus. It is the αβ whole 95057-678 Deoxynucleotide Mix [10mm], 0.5ml 5μmole enzyme with a molecular weight of 157kDa. The enzyme is highly 95057-676 Deoxynucleotide Mix [10mm], 5 x 0.5ml 25μmole pure and does not have contamination of nucleases. The optimal temperature is 41~45°C but with activities at 37~55°C. If Tobacco Vein Mottling Virus (about 10 kb) is used as the template, AMV RTase PCR Clean Up can reverse transcribe 60% of the template, among which 25% are ———————————————————————————— full-length cDNA. Unlike M-MLV reverse transcriptase, AMV reverse Spin-OUT™ for PCR transcriptase can still synthesize cDNA in the presence of 4 mM ———————————————————————————— sodium pyrophosphate, but the optimal temperature has changed to Remove contaminants after PCR 37~40°C. Unit Definition Spin format desalting columns that have the ability to remove One unit is defined as the amount of enzyme required to salts, radioisotpes, dyes, primers and deoxynucleotides (dNTPs) incorporate 1nmol of dTTP into acid-precipitable material in 10 min for nucleic acids following PCR. Two sizes are available to remove at 37°C using poly(A)+ RNA and oligo(dT) as template/primer. contaminating products from PCR products, including <20bp primers, 18 dNTPs and salts or from <32bp primers, dNTPs and salts. Supplied with Enzyme ™ Spin-OUT PCR is for the cleaning of PCR products. PCR-20 • 5× Reaction Buffer: 250mM Tris-HCl, 250mM KCl, 50mM MgCl2, removes contaminating products from PCR products, including 2.5mM spermidine, 50 mM DTT, pH8.3. <20bp primers, dNTPs and salts. PCR-32 removes PCR products Storage Condition from <32bp primers, dNTPs and salts. 200 mM potassium phosphate (pH7.2), 2 mM DTT, 0.2% Triton® FEATURES X-100, 50% Glycerol. • Available for <20bp or <32bp primers Application • Spin format for rapid purification • RT-PCR APPLICATIONS • Real time RT-PCR • Remove primers, dNTPs and other decontaminants from PCR • cDNA library reactions • SAGE • 3' or 5' RACE Cat. No. Description Size 82022-588 Spin-OUT™ PCR-20 10 columns Cat. No. Description Size 82022-590 Spin-OUT™ PCR-32 10 columns 71003-286 AMV Reverse Transcriptase 500U

For further details, visit VWR.com 15 PCR/ RT-PCR RT-PCR Kits A selection of RNA products for the isolation of RNA and the detection, ———————————————————————————— decontamination and digestion of RNase. First-strand cDNA Synthesis Kit Tri-Xtract™ ———————————————————————————— ———————————————————————————— A complete system for efficient first-strand cDNA synthesis For RNA free of protein and DNA contamination ™ from total RNA or mRNA. This Kit contains oligo(dT)18 and random Tri-Xtract is a convenient, ready-to-use reagent designed

nonamer primers (dN)9 according to different experimental needs. for the isolation of total RNA that is free from protein and DNA The kit uses M-MLV Reverse Transcriptase, which is the protein contamination. The isolated RNA is suitable for Northern blots, dot product of mouse leukemia virus gene pol with a single 71 kDa blot hybridization, in-vitro translation, RNase protection assays and + peptide chain and exhibits low RNase H activity. Oligo(dT)18 is poly (A ) selection. designed to prime selectively on mRNA with a poly(A)+ tail. A random Tri-Xtract™ is a monophasic solution of phenol and guanidine nonamer can be used for the generation of cDNA libraries, to copy thiocyanate that maintains the integrity of the RNA during cell mRNAs that do not contain a poly(A)+ tail, or for the synthesis of the disruption and homogenization, the addition of chloroform results in 5' regions of mRNAs. the separation of the homogenate into aqueous and organic phases. Application RNA partitions to the aqueous phase, DNA to the interphase, and • First strand cDNA synthesis for RT-PCR proteins to the organic phase. The total RNA is recovered by isopropyl • Construction of cDNA libraries alcohol precipitation. The DNA and proteins can also be recovered by • Generation of probes for hybridization sequential precipitation from the organic phase.

Cat. No. Description Size 71003-288 First-strand cDNA Synthesis Kit 25 reactions 1-Step RT-PCR Kit ———————————————————————————— Designed for optimal convenience in carrying out highly sensitive and specific RT-PCR in a single tube. 1-Step RT-PCR is a variation on standard (two-step) RT-PCR in which all components of the RT and PCR are mixed in one tube prior to starting the reactions and in which RT and PCR are thus carried out sequentially in one tube. This approach offers tremendous convenience when applied to analysis of single targets from multiple samples of RNA. It also minimizes the possibility for introduction of contaminants into reactions between the RT and PCR steps, since the RT and PCR reactions are carried out sequentially, without opening of reaction tubes between steps. Composed of high quality reagents. The AMV reverse transcriptase is used for RT, and the optimized Taq polymerase is used for PCR. A unique buffer, 10×RT-PCR buffer suitable for both the RT and PCR steps and crucial for successful one-step RT-PCR, is included, along with dNTPs mixture, DEPC-treated water and RNasin. Application ™ • Fast and easy one-tube setup Figure 29: Tri-Xtract scheme. • One-step RT-PCR of any RNA template Tri-Xtract™ is suitable for the isolation of total RNA, including mRNA, hnRNA, ribosomal RNA and tRNA, from small quantities of Cat. No. Description Size tissues (50-100mg) and cells (5 x 106) from human, animal, plant, 71003-290 1-Step RT-PCR Kit 25 reactions yeast, bacterial and viral origin. Total RNA is isolated in under an hour and the subsequent isolation of DNA and protein in less than 2-Step RT-PCR Kit three hours. ———————————————————————————— FEATURES The 2-Step RT-PCR Kit has all the reagents needed to synthesize • Isolate high quality RNA the first-strand cDNA followed by amplification of the cDNA product • Immediate inhibition of RNase using PCR. Oligo(dT)18 and random nonamer primers (dN)9 are • One hour protocol provided according to different experimental needs. The kit uses • Scalable M-MLV Reverse Transcriptase, which is the protein product of mouse • Ability to sequentially isolate RNA, DNA then protein leukemia virus gene pol with a single 71 kDa peptide chain. The APPLICATIONS enzyme has a low RNase H activity. Using oligo(dT)18 or random nonamer as the primer, M-MLV RTase synthesizes the first-strand • RNA suitable for Northern blots, dot blot hybridization, in-vitro + cDNA. PCR reagents provided in this kit are then used to amplify the translation, RNase protection assays and poly (A ) selection cDNA. This kit provides a complete solution from RNA to PCR products. Cat. No. Description Size Application 89131-004 Tri-Xtract™ 100ml • RT-PCR of any RNA template 89131-006 Tri-Xtract™ 200ml • Detecting multiple messages from a single RNA sample

Cat. No. Description Size 71003-292 2-Step RT-PCR Kit 25 reactions 16 For further details, visit VWR.com RNA Purification Arrest™ Extraction Buffer TOTAL Arrest™ ———————————————————————————— ———————————————————————————— A chaotropic RNA extraction buffer Isolation of RNA free of protein & DNA An optimized combination of various chaotropic agents and contamination for RT-PCR ™ RNase inhibitors, which inhibits RNase in 5-10 seconds. Arrest The TOTAL Arrest™ RNA kit isolates RNA from contaminating Extraction Buffer may be used in conjunction with any RNA extraction DNA, proteins, and nucleases using our proprietary binding matrix, method, including extractions based on phenol, chloroform and other pinkRESIN™. The absence of phenol/chloroform extractions make organic solvents and detergents. the Total Arrest™ RNA kit one of the safest methods for isolating The quick 10 minute single step protocol isolates high quality and purifying high quality RNA. The 60 minute protocol (micro kit) total RNA from most species and tissues. Protocol involves is simple; after homogenization, RNA is bound to pinkRESIN™ and ™ homogenization and lysis of samples in Arrest Buffer. After removing washed. The protocol provides an option to remove contaminating cellular debris, pure RNA is precipitated from the supernatant with DNA with a single DNase treatment. Finally RNA is eluted from ethanol. Conveniently packaged as two 50ml bottles for isolation pinkRESIN™. The eluted RNA is ready for further applications, from up to 10 grams of tissue. including Northern and dot blots, reverse transcription or RNase FEATURES protective assays. The kit is supplied in two formats, the Micro for 50 • Rapidly inhibit destructive RNases preps (10-50mg tissue/ prep) and Large for 10 preps (100-500mg • Strong chaotropic buffer for rapid RNA release tissue/ prep). • Compatible with numerous species and tissues Three step protocol: • Suitable for 200 x 50mg extractions 1. Arrest™ Extraction Buffer lyses samples instantly destroying RNase activity. Cat. No. Description Size 2. pinkRESIN™ captures RNA. ™ 82022-524 Arrest Extraction Buffer 200 preps 3. After a brief washing step, pure and protein free RNA is eluted. GET™ Total RNA ———————————————————————————— Small scale, spin format RNA purification The GET™ Total RNA kit isolates total RNA from contaminating DNA, proteins, and nucleases using our GET™ RNA Spin Columns. The <60-minute protocol is simple; after homogenization, RNA is bound to the GET™ RNA Spin Columns and washed. The protocol provides an option to remove contaminating DNA with a single DNase treatment. Finally RNA is eluted from the column. The eluted RNA is ready for any procedure including Northern/slot/dot blots, reverse transcription or RNase protective assays. Three step protocol: Figure 31: TotalARREST™ scheme. 1. Arrest™ Extraction Buffer lyses samples instantly destroying RNase activity. FEATURES 2. GET™ RNA Spin Columns captures RNA. • Rapidly inhibit destructive RNases 3. After a brief wash step, protein free RNA is eluted. • Strong chaotropic buffer for rapid RNA release • High affinity RNA binding matrix to capture RNA • Optional DNase treatment • Compatible with numerous species and tissues • Suitable for animal and plant tissues, cultured cells, blood and bacteria • Suitable for 50 x 50mg (Micro) or 10 x 500mg (Large) extractions APPLICATIONS • For the extraction and purification of protein and DNA free RNA

Cat. No. Description Size 82022-520 Total Arrest™ RNA, micro 50 preps Figure 30: GET™ Total RNA scheme. 82022-522 Total Arrest™ RNA, medi 10 preps FEATURES • Rapidly inhibit destructive RNases • Strong chaotropic buffer for rapid RNA release • High affinity RNA binding spin columns • Optional DNase treatment • Compatible with numerous species and tissues • Suitable for 50 x 50mg extractions APPLICATIONS • For the extraction & purification of protein & DNA free RNA

Cat. No. Description Size 95057-558 GET™ Total RNA 50 preps

For further details, visit VWR.com 17 RNA Purification RNase-DETECT™ NUCLEIC dotMETRIC™ Assay ———————————————————————————— ———————————————————————————— For the detection of RNase contamination A 1μl assay for rapid estimation of DNA, RNA and RNase-DETECT™ is a highly reliable and sensitive method to detect oligo concentrations RNase contamination, which does not utilize unreliable, tedious and The system uses a unique combination of proprietary expensive test strips or radioactive methods. reagents and test strips and is a cost-effective alternative to UV ™ With RNase-DETECT , 10µl of test solution is added to our spectrophotometry as no expensive equipment or cuvettes are calibrated RNA substrate vial, incubated, and the result viewed after necessary. NUCLEIC dotMETRIC™ uses as little as a microliter of your 10 minutes by agarose electrophoresis. sample and provides permanent results in minutes. Samples in the range of 10µg/µl to 1ng/µl can be measured accurately regardless of the isolation method or storage buffer. Oligonucleotides as short as 16 bases have successfully been measured with NUCLEIC dotMETRIC™. Measurements are not affected by protein contamination in the sample. The lower detection limit of the system is 1-2ng/µl. Figure 32: The detection of RNase contamination with RNase-DETECT™. Samples are diluted with dilution buffer and 1-5µl spotted onto The two test samples were compared to the control, showing varying degrees of RNA decontamination. the NUCLEIC Test Strip, which are developed with NUCLEIC Dye in under 2 minutes. The dotMETRIC™ scale and spot diameter are FEATURES compared for accurate nucleic acid concentrations. • Contains aliquots of calibrated proprietary RNase substrate for For increased reproducibility and test reliability the kits are detection of femtogram levels of RNase contamination supplied with an optional Spot Application Device. This allows • Serial Dilution Ladder (SDL) detection technique supplied for rapid, application of samples using fixed volume capillary tips, simplifying clear detection of contamination the application of the nucleic acid solution and improving the • Compatible with a wide variety of reagent solutions, including SDS, reliability of results. Tris, salts, agarose, and so forth The assay is unaffected by the presence of common laboratory • Suitable to assay solutions that can not be treated with DEPC agents, such as reducing agents, chelating agents, detergents, • Supplied with RNase substrate vials, reaction-loading dyes, and amines, sugars, chaotropes, salts and other common laboratory easy to follow protocol for a quick analysis of the results agents. APPLICATIONS Figure 33: NUCLEIC dotMETRIC™ scheme. • For the detection of RNase contamination in a variety of solutions FEATURES Cat. No. Description Size • Sensitive, with lower detection 82022-508 RNase-DETECT™ 50 tests limit of 1-5ng/μl nucleic acid 82022-510 RNase-DETECT™ 100 tests • Only 1µl sample required • Rapid 3 minute assay RNaseOUT™ • Suitable for 300 assays ———————————————————————————— APPLICATIONS Kills RNase on contact • Estimation of DNA, RNA & RNaseOUT™ is uniquely formulated to destroy and remove RNases oligonucleotide concentrations on contact, simply spray and rinse. RNaseOUT™ is non-toxic and • Assay for precious samples, residue free. RNaseOUT™ allows common equipment to be safely and requires only 1µl sample confidently used for all RNA work. ACCESSORIES ™ RNaseOUT is supplied in 250ml easy to apply spray bottles, • Spot Application Device: simply spray on items or area to decontaminate and rinse. Spraying Simplifies the application of prevents waste and is designed to uniformly cover the application samples area. • 1µl Application glass capillary For economy, refilling spray bottles, or cleaning large items, tips ™ RNaseOUT is also available in 1 liter refill bottles. • 1-10µl Sample Application (pipette) Tips for standard pipettes Cat. No. Description Size • Developing Trays 82021-424 RNaseOUT™ 250ml Spray CITED References 82021-426 RNaseOUT™ 1L Refill Akopiants, K. et al (2005) J. Ind. Microbiol. Biotechnol. 33: 141 Nakashima, A., et al (2002) J. Biochem. Soc. 131: 391 Weghofer, M. et al (2001) Ann. Hematol. 80: 733 RNasin Campbell, T. B et al (2000) AIDS. 14:2109 ———————————————————————————— Cat. No. Description Size RNase Inhibitor 82021-416 NUCLEIC dotMETRIC™ >300 assays RNasin is a ribonuclease inhibitor extracted from human placenta 82021-418 NUCLEIC dotMETRIC™ w th Spot Application Device >300 assays with a molecular weight 51kDa. It inhibits the activity of RNase by 82021-420 dotMETRIC™ Spot Application Device 1 specifically binding up to RNase with a non-covalent bond. RNasin, 82021-372 1µl Application Glass Capillary Tips 100 free of RNase or Nickase, can maintain its activity at pH from 5 to 8, 82021-374 Developing Trays 2 and the highest one at pH7.8. Concentration is 20~40 units/µl. 82021-422 Sample Application (pipette) Tips 96 tips Cat. No. Description Size 71003-294 RNasin [20-40U/µl] 1,000 units

18 For further details, visit VWR.com Nucleic Acid Electrophoresis Geno-ElectroPhore™ Power Supplies ———————————————————————————— ———————————————————————————— A mini horizontal gel electrophoresis system ™ A simple and economical device for a variety of agarose GT 250 Power Pack electrophoresis applications, supplied with gel casting tray, combs ———————————————————————————— and horizontal tank. The Complete-Electrophore™ is also supplied Programmable power supply for electrophoresis with a mini power supply. A versatile microprocessor, programmable power supply unit for The ultimate in simplicity and convenience in casting agarose vertical and horizontal gel electrophoresis. gels, light-weight and simple to use. The gel casting tray is designed FEATURES to eliminate the leak problems common with other gel casting trays. • Compact and stackable unit The device allows casting of agarose gels in two sizes: two 5 x 8.3cm • Power capacity: 250 volts, 400mA and 100 watts mini gels and one 10.5 x 8.3cm gel. • Constant voltage or constant current The casting trays are provided with background black strips for • 10 programmable methods available high well visibility. Supplied with two mini gel trays, one large tray and • Timer with alarm function two different size combs. The combs provide either 13 and 6 wells • Safety features: No load detection, shrouded plugs and sockets (0.6 x 0.1cm well) or 17 and 8 wells (0.4 x 0.1cm well) for the large • Suitable for 100-240V and mini gels, respectively. Additional Gel Casting sets that include APPLICATIONS the casting tray, two mini gel trays, one large tray and two different • Programmable power supply for horizontal agarose or vertical size combs are also available. polyacrylamide electrophoresis systems Interlocked ventilated fog free lid allows full view of gel during electrophoresis. Cat. No. Description Size Complete-Electrophore™ 82021-456 GT 250 Power Pack™ 1 Supplied with a power supply unit in addition to Geno- 82021-468 Additional power cord 1 Electrophore™. The power supply has an output of 50 or 100 volt for electrophoresis, which is suitable for most agarose electrophoresis E-Power™ 150 Power Supply applications. ———————————————————————————— Six fixed voltage settings up to 150 volts A micro processor-controlled mini power supply with a small footprint provides the necessary power to run horizontal agarose or vertical polyacrylamide electrophoresis systems. Easy operation, the ability to run two electrophoresis experiments at once, 6 constant voltages, and compact size are the differentiating features. FEATURES • 6 constant voltages: 25, 50, 75, 100, 125 & 150V • Two outlets • Compact size: 115 x 150 x 80mm (W x L x H ); ~0.6kg • Power capacity: 60W, 600mA, 150V • Timer with alarm function • Safety features: No load detection, shrouded plugs and sockets • Suitable for 100-240V Figure 34: Complete-Electrophore™. Cat. No. Description Size FEATURES 89131-014 E-Power™ 150 Power Supply 1 • Eliminates gel leaking issues, no taping required • Cast two gel sizes; two 5 x 8.3cm and one 10.5 x 8.3cm gels E-Power™ 100 Power Supply • Designed for easy well identification ———————————————————————————— • Two combs provided for small and large wells Two fixed voltage settings of 50 & 100 volts APPLICATIONS This power supply is specifically designed for DNA and RNA • A complete horizontal agarose electrophoresis system electrophoresis and allows for two constant voltages of either 50V or 100V. The low cost, light weight and small foot print make it perfect Cat. No. Description Size for teaching labs as well as routine electrophoresis use. 82021-460 Geno-Electrophore™ 1 82021-462 Complete-Electrophore™, MT-108 with power supply 1 FEATURES 95057-528 E-Power 100 Power Supply 1 • 50V or 100V constant voltage • Power capacity: 40W, 400mA, 100V 82021-464 Comb for 1 x 4mm wells 1 • Safety features: No load detection, shrouded plugs and sockets 82021-466 Comb for 1 x 6mm wells 1 • Compact & lightweight: 60 x 40 x 120 mm (W x L x H); ~0.3kg 82021-470 Large gel tray (10.5 x 8.3cm) 1 • Developed specifically for DNA and RNA electrophoresis 82021-472 Mini gel tray (5.0 x 8.3cm) 1 • Suitable for 100-120V 89168-112 Casting Tray 1 Gel Casting Set Cat. No. Description Size 71003-312 1 (Casting tray, 2 mini gel trays, large tray & 2 combs) 95057-528 E-Power™ 100 Power Supply 1

For further details, visit VWR.com 19 Nucleic Acid Electrophoresis G-CAPSULE™ GET™ AGAROSE DNA ———————————————————————————— ———————————————————————————— Electroelution device for the rapid purification of Rapid purification of DNA from agarose gels nucleic acids from electrophoresis gels This kit is based on our GET™ Spin Columns, spin columns with a Electroelution of nucleic acids and proteins has many advantages high binding affinity for DNA. as it avoids centrifugation, vortexing, heating, precipitation and GET™ AGAROSE DNA method involves release of DNA fragments allows minimal manipulation of samples. Electroelution normally from gel pieces followed by capture of nucleic acids on the GET™ Spin involves dialysis tubing, which results in extreme dilution of precious Columns, washing, and elution of the clean nucleic acid fragments in samples. G-Capsule™ is a simple electroelution device that excises a suitable buffer. DNA or protein bands and elutes your sample in a final volume of The GET™ AGAROSE DNA kits are supplied with enough reagents ~30µl. for 50 or 100 preps. G-CAPSULE™ has two parts, G-Pick™ and G-Trap™. The user simply picks up the protein or nucleic acid band with the G-Pick™ and assembles it with the G-Trap™. The assembled G-CAPSULE™ is submerged in electrophoresis buffer on a horizontal electrophoresis system and the protein or nucleic acid is rapidly eluted into the G-Trap™.

Figure 37: GET™ AGAROSE DNA scheme. FEATURES • Extract DNA fragments from agarose gels • Rapid DNA isolation • DNA ready for downstream applications, including ligations • Suitable for 100-20,000bp fragments • Compatible with TAE and TBE buffer gels

Figure 35: A schematic of the G-CAPSULE™ procedure. APPLICATIONS • Isolation of DNA fragments from agarose gels FEATURES • Cleaning and removing of contaminants from DNA samples • Rapid electroelution of nucleic acids and proteins Cat. No. Description Size • Sample recovered in a small volume (25-50μl) ™ • Recovery is as high as 90% 95057-574 GET AGAROSE DNA 50 preps 95057-576 GET™ AGAROSE DNA 100 preps APPLICATIONS • Extraction of >20bp DNA and RNA or for >4kDa proteins geneEXIT™ ACCESSORIES ———————————————————————————— • G-CAPSULE™ Weight: A small weight device that prevents Isolation of nucleic acids from agarose gels G-CAPSULE™ from floating during electroelution The kit is based on pinkRESIN™, a high capacity, proprietary binding resin matrix for nucleic acids. pinkRESIN™ has an enhanced binding affinity for DNA and RNA and resuspends with ease, thus eliminating loss and damage of nucleic acids, a frequent problem with other binding resin matrices for DNA and RNA isolation. pinkRESIN™ is a non-toxic matrix. geneEXIT™ method involves release of nucleic acid fragments from gel pieces followed by capture of nucleic acids with pinkRESIN™, washing, and elution of the clean nucleic acid fragments in a suitable buffer. The geneEXIT™ method can be carried out with or without the use of spin columns; however the use of spin columns simplifies the Figure 36: The G-CAPSULE™ weight. protocols. Cited References FEATURES Chatterjee, S et al (2012) Acta Biochim Biophys Sin. 259:68 • Rapid nucleic acid isolation Chatterjee, S et al (2012) Acta Biochim Biophys Sin. 44:259 • DNA & RNA ready for further applications, including ligations Chatterjee, S et al (2012) Acta Biochim Biophys Sin. 10:1093. • Suitable for 100-20,000bp fragments Cardi, D et al (2010) J. Biol. Chem. 285:26406-26416. Crosslin, James (2009) HortScience. 44: 1790 - 1791 • Compatible with TAE and TBE buffer gels Li, X. et al (2004) Euro J of Phycology. 39: 1, 73-82 • Supplied with or without spin columns Beeson, K. et al (2002) Microbiology. 148: 179 - 189 Yeager, M. et al (2001) Circ. Res. 88: 2e - 11 APPLICATIONS Robu, M. et al (2001) PNAS. 98: 8211 • Isolation of DNA/RNA fragments from agarose gels Dudley, E. et al (2001) Microbiology. 147: 215 - 224 • Cleaning contaminants from DNA & RNA samples Brezinschek, Hans-Peter et al (2000) Int. Immunol. 12: 767 Tanaka, K. et al (1999) Yeast. Vol 15: 11: 1133-39 Cat. No. Description Size ™ Cat. No. Description Size 82022-730 geneEXIT 100 preps ™ 82021-228 G-CAPSULE™ 55/box 82022-732 geneEXIT with spin columns 100 preps 82021-230 G-CAPSULE™ Weight 1 weight

20 For further details, visit VWR.com Nucleic Acid Electrophoresis ™ The following table highlights the different dyes and SDS present Glow Dyes ™ ———————————————————————————— in the loading dyes, in addition, the Glow Dyes are also available DNA/RNA loading dyes with premixed ethidium without ethidium bromide (Universal Dyes). All dyes are supplied in bromide to reduce exposure hazards 1.5ml vials. Nucleic acids are routinely visualized on agarose gels with the highly toxic ethidium bromide stain. High concentrations of ethidium bromide are either added to the molten agarose prior to pouring or to a large volume of staining buffer to stain the nucleic acids after electrophoresis. Either method results in a large amount of ethidium bromide to handle and dispose of. Catalog Description The Glow™ Dyes are designed to minimize the use of ethidium Blue Bromophenol Cyanol Xylene SDS Bromide Ethidium DNA Glow™ Dyes [6X] bromide and the risk associated with the regular use of ethidium ™ bromide. Glow™ Dyes have been specifically formulated for nucleic 82022-468 Glow BromoBlue Dye Yes No No Yes ™ acid electrophoresis and contain an optimized concentration of 82022-470 Glow CyanoBlue Dye Yes Yes No Yes buffer agents, tracking dyes and ethidium bromide. These loading 82022-472 Glow™ CleanAway™ Dye Yes Yes Yes Yes dyes reduce exposure and many of the problems associated with DNA Universal Dyes [6X] ethidium bromide use and disposal. Simply add an appropriate 82022-462 BromoBlue Universal Dye Yes No No No volume of Glow™ Dyes to your sample, load the gel and then visualize 82022-464 CyanoBlue Universal Dye Yes Yes No No with UV; no need for additional ethidium bromide staining or addition 82022-466 CleanAway™ Universal Dye Yes Yes Yes No of ethidium bromide to the agarose. RNA Glow™ Dyes [2X] ™ The DNA Glow Dyes are Ficoll based and are supplied at a 6X 82022-476 Glow™ RNA Dye Yes Yes No Yes ™ concentration and the RNA Glow Dyes are formamide based and are RNA Universal Dyes™ [2X] at a 2X concentration. 82022-474 Universal RNA Dye Yes Yes No No The DNA and RNA Loading Dyes are offered in multiple formats to meet all your needs and have variations in the following components: Dyes: The loading dyes use Bromophenol Blue that migrates at ~300bp (0.7-1.7% agarose) or ~100bp (2.5-3.0% agarose) to highlight the migration front and Xylene Cyanol FF (CyanoBlue™ Loading Dyes) that migrates at ~4,000bp (0.7-1.7% agarose) or ~800bp (2.5-3.0% agarose) to help resolve higher molecular weight nucleic acids. SDS: The detergent SDS (sodium dodecyl sulfate) is supplied in the CleanAway™ Loading Dyes and these are recommended for use with DNA that has a high level of DNA binding proteins. The SDS eliminates DNA-protein interactions, which prevents poor DNA migration, DNA sticking to the wells or band shifts. In addition, SDS treatment prevents long cohesive ends reannealing and Figure 38: DNA/RNA loading dyes producing artifactual DNA bands. Ethidium Bromide: The benefits of ethidium bromide is the Glow™ Loading Dyes is highlighted above, however for researchers who want the benefits of our loading dyes without the ethidium bromide, we offer our Universal Loading Dyes FEATURES • Provide intense DNA/RNA bands with little background or band distortion • Compatible with all agarose or acrylamide gel electrophoresis. • Glow™ RNA Dye is ideal for quick screenings of RNA preps • There is no need to use toxic formaldehyde agarose gels for checking RNA integrity with Glow™ RNA Dye • Also available are Universal Loading Dyes without ethidium bromide and containing different tracking dyes and the option of added SDS APPLICATIONS • Suitable as sample loading dye for electrophoresis application • Dye for tracking electrophoresis migration and visualization of nucleic acids

For further details, visit VWR.com 21 Nucleic Acid Electrophoresis DNA Ladders DNAmark™ 100bp Ladder ———————————————————————————— ———————————————————————————— ™ Consists of 9 fragments ranging from 100-1500 base pairs (bp). DNAmark Logic DNA Ladder This molecular weight marker is specially designed for identifying ———————————————————————————— small DNA fragments and allowing for quantitative estimation of the Contains 16 DNA bands ranging from 100bp to 10Kb, formulated DNA fragments in the gel at the same time. so that each band contains an amount of DNA that correlates logically to its size, allowing the user to estimate both the size and the quantity of specific fragments at a glance. It is particularly useful for protocols such as probe labeling, DNA sequencing and optimizing Base Pairs DNA Mass (ng) insert/vector ratio in ligation reactions. 1,500 150 1,000 100 Base Pairs DNA Mass (ng) 800 80 10,000 100 600 60 8,000 80 500 50 6,000 60 400 40 5,000 50 300 30 4,000 40 200 20 3,000 30 100 10 2,000 20 Figure 41: DNAmark™ 100bp DNA Ladder on a 1% TAE agarose gel, 1,500 150 stained with LabSafe™ Nucleic Acid Stain. 1,000 100 Cat. No. Description Size 800 80 89233-912 DNAmark™ 100bp Ladder 100 loads 600 60 500 50 400 40 300 30 200 20 100 10 Figure 39: DNAmark™ Logic DNA Ladder on a 1% TAE agarose gel, stained with LabSafe™ Nucleic Acid Stain.

Cat. No. Description Size 89233-910 DNAmark™ Logic DNA Ladder 100 loads

DNAmark™ 100bp Plus Ladder ———————————————————————————— Consists of 13 DNA fragments ranging in size from 100-3000 base pairs (bp). 6µl will yield at least 30ng DNA in any single band. The intensity of the 500bp and 1500bp bands has been increased to serve as a reference for easy identification.

Base Pairs 3,000 2,000 1,500 1,000 900 800 700 600 500 400 300 200 100 Figure 40: DNAmark™ 100bp Plus DNA Ladder on a 1% TAE agarose gel, stained with LabSafe™ Nucleic Acid Stain.

Cat. No. Description Size 89233-914 DNAmark™ 100bp Plus Ladder 100 loads 22 For further details, visit VWR.com Nucleic Acid Electrophoresis Accessories, Buffers & Chemicals Ethidium Bromide ———————————————————————————— ———————————————————————————— Cat. No. Description Size Electrophoresis Running Buffers 82023-294 Ethidium Bromide Solution [10mg/ml] 10ml ———————————————————————————— 89168-116 Ethidium Bromide Solution [0.625mg/ml] 5ml Molecular grade, concentrated running buffers for DNA and RNA 71003-400 Ethidium Bromide 5g electrophoresis. ™ Cat. No. Description Size FASTsilver 89167-720 TAE [50X] 0.5L ———————————————————————————— 82021-492 TAE [50X] 1L A rapid silver stain for nucleic acids 82021-494 TAE [50X] 1gal A nanogram sensitive silver staining kit that produces crystal clear 82021-496 TBE [10X] 1L background and maximal sensitivity needed for critical analysis. 82021-498 TBE [10X] 1gal A unique formulation of reagents that leaves the background ™ 95029-252 TBE-Urea Sample Buffer 30ml clear and produces sharp images of nucleic acid bands. FASTsilver 82021-508 MOPS Running Buffer [10X] 1L detects as little as 0.3ng nucleic acid. The kit contains ready to use reagents for 25 mini-gels and comes with a simple to follow 60-90 82021-510 MOPS Running Buffer [10X] 1gal minute protocol. 95043-526 MOPS/SDS Running Buffer [20X] 0.5L FEATURES Agarose Powders • Stains both nucleic acids and proteins ———————————————————————————— • Produces clear background for maximum visibility Biotechnology grade agarose powders • Protocol time 60-90 minutes APPLICATIONS Cat. No. Description Size • Staining of nucleic acids and proteins in electrophoresis gels 71003-518 Agarose I (All purpose agarose) 5g Cited References 95057-706 Agarose I (All purpose agarose) 25g Reed, D. et al (2005) Plant Cell Reports. 24:15-24 95057-708 Agarose I (All purpose agarose) 100g Ortega, N. et al (2005) Mol. Biol. Cell 16: 3028 Venkatesh, S.G. et al (2004) Am. J. Physiol. Cell Physiol. 286: C365 95057-714 Agarose I (All purpose agarose) 500g Melody, J. L. et al (2004) J. Anim. Sci. 82: 1195 95057-710 Agarose II (Low melting agarose) 25g Delcroix, J. D. et al (2003) Neuron 39: 69 95057-712 Agarose II (Low melting agarose) 100g Wu, C. et al (2003) Am. J. Respir. Cell Mol. Biol. 2: 731 Kralj, S. et al (2002) Appl. Envir. Microbiol. 68: 4283 71003-332 Agarose III (Pulse field gel electrophoresis) 25g 71003-334 Agarose III (Pulse field gel electrophoresis) 100g Cat. No. Description Size 71003-336 Agarose IV (Highest gel strength) 25g 82021-392 FASTsilver™ 25 mini gels 71003-338 Agarose IV (Highest gel strength) 100g GELPLATE-clean™ LabSafe™ Nucleic Acid Stain ———————————————————————————— ———————————————————————————— Spray and wipe; specifically formulated for LabSafe™ Nucleic Acid Stain is a new and safe nucleic acid electrophoresis gel plates stain for the visualization of double-stranded DNA, single-stranded ™ DNA, and RNA in agarose gels. The dyes are developed to replace Clean your electrophoresis plates with GELPLATE-clean and toxic Ethidium Bromide (a potent mutagen), commonly used in gel prevent poor electrophoretic band migration, band distortions, and poor image development. Simply spray and wipe clean your electrophoresis for visualization of nucleic acids in agarose gels. ™ LabSafe™ Nucleic Acid Stain is non-carcinogenic by the Ames-test. electrophoresis plates with GELPLATE-clean . The results are negative in both the mouse marrow chromophilous FEATURES erythrocyte micronucleus and mouse primary spermatocyte • Suitable for sequencing and protein electrophoresis gel plates chromosomal aberration tests. • Removes proteins, nucleic acids, fats, lipids, radioisotopes and LabSafe™ Nucleic Acid Stain emits green fluorescence when other contaminants from electrophoresis gel plates bound to dsDNA and red fluorescence when bound to ssDNA or RNA. • Supplied in 2 x 250ml easy to apply spray bottles It has two excitation wavelength peaks when bound to nucleic acid, • Refill solutions are offered separately at 290nm and 490nm. APPLICATIONS • Cleans sequencing and electrophoresis gel plates

Cat. No. Description Size 82021-306 GELPLATE-clean™ Spray 2 x 250ml 82021-310 GELPLATE-clean™ Refill 2 x 1L ™ Figure 42: Plasmid DNA stained with LabSafe Nucleic Acid Stain 82021-308 GELPLATE-clean™ Refill for Solution I 1L 95029-166 GELPLATE-clean™ Refill for Solution II 1L Cat. No. Description Size 89233-908 LabSafe™ Nucleic Acid Stain [10,000X] 1ml

For further details, visit VWR.com 23 Hybridization Buffers EKONO™ ———————————————————————————— For cleaner backgrounds A high performance hybridization buffer system providing researchers with unprecedented simplicity, reliability, accuracy, and cost saving for hybridization. Protocol involves incubation of blots with denatured probes in EKONO™ hybridization buffer followed by washing of the blot to remove excess unhybridized probes. Easy-to-use EKONO™ provides high resolution and sensitivity. Suitable for Southern/Northern or Southwestern hybridization analysis, hybridization with oligonucleotides or other types of probes, and for hybridization screening of gene libraries. CITED REFERENCES Guo, H. et al (2010) Antimicrob. Agents Chemother. doi:10.1128/AAC.00989 Guo, H. et al (2007) J. Virology 81: 10072 Guo, H. et al (2007) J. Virology 81: 12472 Dougherty, A. M. et al (2007) Antimicrob. Agents Chemother. 51:4427 Hodson, J. et al (2003). Nuc. Acid Res. e31: e134 Kim, Mee-Jung et al (2002) PNAS. 99: 10096 Moraleda, G. and Taylor, J. (2001) J. Virol. 75: 10161

Cat. No. Description Size 82021-338 EKONO™ 1L HYB-LINK™ ———————————————————————————— Formamide-free hybridization buffer Hyb-LINK™ is a high performance formamide free hybridization solution for Northern, Southern and Dot Blots, which increases sensitivity of random-primer DNA probes. Hyb-LINK™ capitalizes on increased sensitivity of blot hybridization without increasing background and therefore provides reliable and accurate hybridization. Hyb-LINK™ is compatible with both RNA and DNA probes.

Cat. No. Description Size 95029-218 Hyb-LINK™ 125ml 95029-220 Hyb-LINK™ 4 x 125ml Other Hybridization Buffers ———————————————————————————— Cat. No. Description Size 89167-768 Church & Gilbert’s [2X] 250ml 89167-770 Church & Gilbert’s [2X] 0.5L 95043-512 Denhardt Solution [100X] 50ml 82021-476 Hybridization Denaturing Solution 1L 82021-478 Hybridization Denaturing Solution 1gal 82021-480 Hybridization Neutralizing Solution 1L 82021-482 Hybridization Neutralizing Solution 1gal 89167-710 SSC [20X] 0.5L 82021-484 SSC [20X] 1L 82021-486 SSC [20X] 1gal 89167-712 SSPE [20X] 0.5L 82021-488 SSPE [20X] 1L 82021-490 SSPE [20X] 1gal 95043-538 TMAC Hyb Solution [3X] 1L 95029-246 TNT Buffer 100ml 95029-248 TNT Buffer 500ml

24 For further details, visit VWR.com Yeast Research Tools EZ Yeast™ Plasmid Prep Fast Yeast Transformation™ ———————————————————————————— ———————————————————————————— A simple yeast plasmid miniprep method For the rapid transformation of yeast The EZ Yeast™ Plasmid Prep kit is designed for the rapid isolation Fast Yeast Transformation™ kit is a rapid single step yeast of 2μ plasmids from yeast patches or yeast grown in small liquid transformation kit that takes less than 10 minutes to prepare cultures. The kit is adapted from the alkaline lysis of E. coli, competent yeast cells. The competent yeast cells can be used by providing modified alkaline lysis buffers and our LongLife™ immediately or frozen for later use. This method is suitable for both Zymolyase®, a highly stable enzyme. The EZ Yeast™ Plasmid Prep kit circular and linear plasmid transformations. The protocol involves makes it possible to isolate high yield yeast plasmid without the use simply suspending yeast cells in the supplied Competent Buffer and of glass beads, phenol, or repeated vortexing. the cells are then ready to receive transforming DNA. Introduce DNA LongLife™ Zymolyase®, a high performance lytic enzyme to the competent cells and incubate for transformation. preparation, efficiently releases plasmids with a yield of up to 0.1- FEATURES 0.3ng for most 2µ-based plasmids from a 1.5ml culture (1 prep). 5 6 ™ • High transformation efficiency: 10 -10 transformants/ μg circular EZ Yeast Plasmid Prep is suitable even for low copy number yeast DNA plasmids. • Broad spectrum: Compatible with C. albicans, S. pombe, P. The kit can be used for plasmid isolation from colonies, patches pastoris, or S. cerevisiae on plate, or liquid culture. The recovered plasmid is in TE buffer and • Transformation procedure takes less than an hour ready for use in any molecular biology application such as E. coli • Frozen competent cells are good for use for up to 6 months transformations, PCR, etc. • Simple protocol for multiple plasmid transformation APPLICATIONS • For the generation of competent yeast cells • Generates plasmids suitable for multiple downstream applications CITED REFERENCES Ito, J. et al (2011) Plant Cell Physiol. 52:539 Yang, J. et al (2011) Mol. Microbiol. 79:872 Kim, J. et al (2011) Plant Cell Physiol. 52:2136 Takano, S. et al. (2010) Plant Cell Physiol. 51:62 Kikis, E. A. et al (2009) PLoS Genet 5(1):e1000352 Aihara, T et al (2009) Biol. Reprod. 80:762 Hayashi, C., et al (2008) J. Biol Chem. 283:14801 Morokuma, Y. et al (2007) J. Biol. Chem. 282:24806 Lin, Y. F. et al (2007) J. Biol. Chem. 282:16783 Ono, Y et al. (2006) J. Biol. Chem.281:18519 Lin, Y. et al. (2006) PNAS. 103:15617 Ueki, N and Hayman, M.J. (2003) J. Biol. Chem. 278:24858 Figure 43: The EZ Yeast™ Plasmid Prep scheme. Uhl, M. et al (2003) EMBO J. 22:2668 Xia, H. et al (2003) Plant Cell 15:449 FEATURES Jia, Huu-Jun et al (2003) Plant Cell. 15:449 Asawatreratanakul, K. et al (2003) Eur. J. Biochem. 270:4671 • Minimize time to analyze yeast plasmids; no need to shuttle Osman, A. et al (2001) J. Biol. Chem. 276:10072 plasmids to E.coli • Yield of 0.1-0.3ng/ 1.5ml culture Cat. No. Description Size ™ • No requirement for glass beads, phenol or excessive vortexing 82023-072 Fast Yeast Transformation 120 preps • Suitable for colonies, patches and liquid culture • Purified plasmid DNA suitable for PCR amplification, Yeast Related Buffers transformations and other downstream applications ———————————————————————————— APPLICATIONS Quality DNase and RNase free reagent solutions. • Extraction of 2μ plasmids from yeast Cat. No. Description Size • Generates plasmids suitable for multiple downstream applications 82023-110 Sorbitol solution [1M] 100ml CITED REFERENCES 82023-112 Lithium acetate [1M] 100ml Baldwin, E. et al (2005) Nuc. Acid Res. 33: 1021 82023-114 Calcium chloride [1M] 100ml Hu, J. et al (2005) Nuc. Acid Res. 33: 3271 Belanger, K. et al (2004) J Biol. Chem. 279: 43530 Sabourin, M. et al (2003) Nuc. Acid Res. 31: 4373 Walikonis, R. S. et al (2001) J. Neurosci. 21: 423

Cat. No. Description Size 82023-264 EZ Yeast™ Plasmid Prep 100 preps

For further details, visit VWR.com 25 Yeast Research Tools Yeast Geno-DNA-Template™ LongLife™ Enzyme Preparations ———————————————————————————— ———————————————————————————— Extraction of high quality genomic DNA from yeast Enzymes regularly used in laboratory applications often require The Yeast Geno-DNA-Template™ extraction kit isolates high quality preparation of fresh solution before each use. Making fresh enzyme genomic DNA from yeast. The kit is based on a two-step lysis of solution for each application is time consuming and wasteful. A wide yeast cells, using LongLife™ Zymolyase® and LongLife™ Proteinase K, variety of enzyme preparations in a ready-to-use format are offered. ™ followed by the removal of proteins and other cellular impurities by LongLife enzyme preparations have a long shelf life and no precipitation and centrifugation. weighing or buffer preparation is needed; simply take an aliquot ™ The clean, genomic DNA is collected by precipitation and the high and add in your sample. LongLife enzyme preparations contain cofactors necessary for optimal enzymatic activity. Supplied in yield of extracted DNA has an average 100kb length and has A260/280 1.8-2.0. suspension form and when stored properly have a one year shelf life. The kit is supplied with reagents for extracting DNA from 100 x ENZYMES OFFERED 1.5ml yeast cultures. • LongLife™ Zymolyase® for the digestion of yeast and fungal cell walls. • LongLife™ Lysozyme for the digestion of bacterial cell walls. • LongLife™ PE LB Lysozyme for the digestion of bacterial cell walls and fully compatible with the PE LB buffer system. Reduces viscosity build-up due to presence of nucleases. • LongLife™ Proteinase K for the digestion of proteins in nucleic acid preparations. • LongLife™ Nuclease for the removal of nucleic acids. • LongLife™ RNase for the digestion of RNA. • LongLife™ DNase for the digestion of DNA.

Figure 44: Yeast Geno-DNA-Template™ scheme. FEATURES • For extraction of genomic DNA from yeast cultures • Compatible with PCR and other downstream applications APPLICATIONS • Designed for the preparation of high yield, genomic DNA template from yeast

Cat. No. Description Size 82021-534 Yeast Geno-DNA-Template™ 100 preps LongLife™ Zymolyase® ———————————————————————————— ™ ® A high performance LongLife Zymolyase preparation is supplied Figure 46: LongLife™ Enzymes are highly stable. Each enzyme preparation in a ready-to-use solution form. The enzyme contains high yeast lytic was tested over a period of 4 weeks at 37°C & compared with LongLife™ activity with low non-specific activity. Suitable for yeast cell lysis, enzyme preparations stored at -20°C. LongLife™Zymolyase® (1.5units/μl) spheroplast preparation, glucan hydrolysis and more. The enzyme was tested with freshly grown yeast suspension by monitoring the decrease in preparation has been stabilized and can be stored at -20°C. absorbance of the suspension. LongLife™ Lysozyme (1500units/μl) was tested by monitoring the decrease in the absorbance of Micrococcus lysodeikticus suspension. LongLife™ Proteinase K (5mg/ml) was assayed with our Protease Assay kit (Cat. No. 82023-260). No measurable loss of activity was noticed. Cited References LongLife™ RNase Tashiro, R. et al (2010) Crop Sci. 50:1260 LongLife™ Lysozyme Butcher, B.G. et al (2011) J. Bacteriol. 193:4598 Ermolova, N. et al (2011) Hum. Mol. Genet. 20:3331 Figure 45: LongLife™ Zymolyase® are highly stable. The enzyme Markel, E. et al (2011) J. Bacteriol. 193:5775 LongLife™ Proteinase K preparation was tested over a period of 4 weeks at 37°C: and compared Whitaker, V. M. et al (2007) J. Amer. Soc. Hort. Sci.132:534 with LongLife™ enzyme preparations stored at -20°C. LongLife™ Zymolyase® (1.5units/μl) was tested with freshly grown yeast suspension Cat. No. Description Size by monitoring the decrease in absorbance of the suspension. 82021-514 LongLife™ Zymolyase® [1.5U/µl] 2 x 0.5ml 82021-516 LongLife™ Lysozyme [1,500U/µl] 2 x 0.5ml CITED REFERENCES 82021-528 LongLife™ PE LB Lysozyme [1,500U/µl] 2 x 0.5ml Gray, P. et al (2006) Mol Cell Prot. 6:514 ™ Saribas, A. et al (2004) Glycobiology 14:1217 82021-518 LongLife Proteinase K [5mg/ml] 2 x 0.5ml 82021-520 LongLife™ Nuclease [10U/µl] 2 x 0.5ml Cat. No. Description Size 82021-524 LongLife™ RNase [10U/µl] 2 x 0.5ml 82021-514 LongLife™ Zymolyase® 2 x 0.5ml 82021-526 LongLife™ DNase 0.5ml 26 For further details, visit VWR.com Buffers & Reagents DNA OUT™ Molecular Grade Water ———————————————————————————— ———————————————————————————— DNA OUT™ is formulated to quickly remove DNA and RNA RNase, DNase & protease free water contaminations from glass and plastic wares. Simply spray and Molecular Grade Water™ is suitable for use in molecular biology ™ rinse. DNA OUT is non-toxic and leaves no residues to interfere with applications that demand a high quality of water and assurance that ™ downstream applications. The fast acting DNA OUT allows laboratory the water is free from DNase, RNase and protease contamination. equipment and work areas to be made free from DNA contamination. No toxic agents, such as DEPC, are used in the manufacturing of Ideal for cleaning microfuge tubes, reaction tubes, PCR machine Molecular Grade Water™, eliminating DEPC interference of enzymatic surface, pipettes, lab benches, lab equipments, and so forth. reactions. Each lot is quality tested for the absence of RNase, DNase ™ DNA OUT is supplied in 250ml easy to apply spray bottles. For and protease contamination. economy, refilling spray bottles, or cleaning large items, DNA OUT™ is Cited References also available in 1 liter containers. Itzek, A. et al (2011) J. Bacteriol. 193:6912 Cat. No. Description Size Block, J. et al (2003) J. Anim. Sci. 81:1590 McKillip, J. et al (1998) Appl. Envir. Microbiol. 64:4264 82021-436 DNA OUT™ 250ml 82021-438 DNA OUT™ 1L Cat. No. Description Size 95043-414 Molecular Grade Water™ 0.5L Molecular Biology Buffers & 95043-416 Molecular Grade Water™ 1L 82021-430 Molecular Grade Water™ 1gal Chemicals 82021-434 Molecular Grade Water™ 4 x 1gal/ case ———————————————————————————— DEPC-Treated Water Molecular Biology Universal Kit ———————————————————————————— ———————————————————————————— RNase free water A selection of 12 x 25ml molecular biology relevant buffers. These buffers are routinely used in molecular biology and are all For all your RNA research needs. Water is treated with DNase and RNase free. The buffers supplied are listed below. Diethylpyrocarbonate (DEPC).

Cat. No. Description Size Cat. No. Description Size ™ 82021-512 Molecular Biology Universal Kit 12 x 25ml 82021-260 DEPC-Treated Water 1L Ammonium acetate [5M] 82021-262 DEPC-Treated Water™ 100ml Calcium chloride [1M] 82021-264 DEPC-Treated Water™ 500ml EDTA, pH8.0 [0.5M] Lithium acetate [1M] Magnesium chloride [1M] Proteomic Grade Water Potassium acetate [3M] ———————————————————————————— SDS [10%] For 2D electrophoresis and mass spectrometry Sodium acetate, pH5.2 [3M] Removes worries of protein and dust contamination and Sodium chloride [5M] TE Buffer [100X] improves quality and reproducibility of 2D electrophoresis and mass Tris, pH7.0 [1M] spectrometry results. Tris, pH8.0 [1M] Cat. No. Description Size 82021-370 Proteomic Grade Water 1L Research Grade Water ———————————————————————————— Endotoxin Free Water ———————————————————————————— Endotoxin, DNA, RNA & protease free water Endotoxin Free Water is free of endotoxins and enzymes, including proteases. Certified tested by the Limulus amebocyte lysate (LAL) test for endotoxins and determined to be <0.0050EU/ml.

Cat. No. Description Size 89167-874 Endotoxin Free Water 0.5L 89167-876 Endotoxin Free Water 1L

For further details, visit VWR.com 27 Buffers & Reagents Buffers & Reagents ———————————————————————————— RNase and DNase free A selection of DNase and RNase free buffers designed to ensure high quality research.

Cat. No. Description Size Cat. No. Description Size 95029-290 Acid Citrate Dextrose (ACD) Solution A 25ml 82021-474 SDS [10%] 100ml 95029-292 Acid Citrate Dextrose (ACD) Solution B 25ml 89167-708 SDS [20%] 1L 71003-330 Agar 1kg 89167-706 SDS [20%] 0.5L 71003-328 Agar 500g 95029-286 SM Buffer 100ml 82023-100 Ammonium Acetate [5M] 100ml 95029-288 SM Buffer 0.5L 71003-354 Ampicillin Sodium Salt 100g 82023-078 SOB Media 50ml 71003-352 Ampicillin Sodium Salt 25g 71003-462 Sodium Acetate 500g 95043-510 BBS [2X] 1L 71003-464 Sodium Acetate 1kg 95043-508 BBS [2X] 0.5L 82023-096 Sodium Acetate, pH5.2 [3M] 100ml 71003-362 Bromophenol Blue (ACS Grade) 25g 95029-192 Sodium Azide Solution [1%] 100ml 71003-364 Bromophenol Blue (ACS Grade) 50g 71003-466 Sodium Bicarbonate 500g 71003-370 Calcium Chloride (Dihydrate) 500g 71003-468 Sodium Bicarbonate 1kg 71003-372 Calcium Chloride (Dihydrate) 1kg 71003-524 Sodium Carbonate (Anhydrous) 500g 71003-376 Carbenicillin Disodium Salt 1g 71003-526 Sodium Carbonate (Anhydrous) 1kg 71003-374 Carbenicillin Disodium Salt 250mg 95029-574 Sodium Chloride 500g 71003-378 Cesium Chloride 100g 71003-470 Sodium Chloride 1kg 71003-380 Cesium Chloride 500g 82023-090 Sodium Chloride Solution [5M] 100ml 89167-780 CTAB Extraction Buffer 60ml 71003-542 Sodium Hydroxide 100g 89167-782 CTAB Extraction Buffer 125ml 82023-092 Sodium Hydroxide Solution [2N] 100ml 71003-388 DEPC (Diethylpyrocarbonate) 5g 71003-472 Sodium Phosphate (Dibasic) 500g 71003-390 DEPC (Diethylpyrocarbonate) 25g 71003-474 Sodium Phosphate (Dibasic) 1kg 89167-784 Gram Negative Lysis Buffer 125ml 71003-476 Sodium Phosphate (Monobasic) 500g 89167-786 Gram Negative Lysis Buffer 250ml 71003-478 Sodium Phosphate (Monobasic) 1kg 71003-190 Gram Negative Lysis Buffer 500ml 82023-110 Sorbitol Solution [1M] 100ml 71003-192 Gram Negative Lysis Buffer 1L 89167-790 STE (TEN) Buffer [10X] 1L 95043-520 Imidazole [2M] pH7.5 1L 89167-788 STE (TEN) Buffer [10X] 0.5L 71003-420 IPTG (Molecular Biology Grade) 10g 95043-532 STET Solution 0.5L 95029-570 IPTG (Molecular Biology Grade) 1g 95043-534 STET-E1 Solution 1L 82023-106 IPTG (Molecular Biology Grade) 5g 95043-536 STM Buffer 1L 82023-112 Lithium Acetate [1M] 100ml 82023-084 TE Buffer [100X] 100ml 71003-430 Magnesium Chloride 100g 89167-718 TE Buffer [10X] 1L 71003-432 Magnesium Chloride 500g 89167-716 TE Buffer [10X] 0.5L 82023-086 Magnesium chloride [1M] 100ml 71003-486 Tricine 100g 95043-522 Magnesium Sulfate Solution [1M] 1 L 71003-488 Tricine 500g 71003-438 MOPS (3-[N-Morpholino] propane-sulfonic acid) 100g 95029-578 Tris Base 500g 71003-440 MOPS (3-[N-Morpholino] propane-sulfonic acid) 250g 71003-490 Tris Base 1kg 89167-776 Phenol Red Solution [0.5%] 100ml 71003-492 Tris HCl 500g 89167-778 Phenol Red Solution [0.5%] 250ml 71003-494 Tris HCl 1kg 82023-098 Potassium Acetate [3M] 100ml 82023-080 Tris, pH7.0 [1M] 100ml 82023-088 Potassium Chloride Solution [1M] 100ml 89167-742 Tris, pH7.2 [1M] 100ml 71003-520 Potassium Chloride, KCl 500g 89167-744 Tris, pH7.5 [1M] 100ml 71003-522 Potassium Chloride, KCl 1kg 82023-082 Tris, pH8.0 [1M] 100ml 82023-094 Potassium Hydroxide Solution [2N] 100ml 71003-102 Tris, pH8.5 [1M] 100ml 71003-538 Potassium Iodide, KI 500g 95029-256 Tris, pH9.0 [1M] 100ml 71003-540 Potassium Iodide, KI 1kg 71003-496 Tryptone 100g 71003-450 Potassium Phosphate (Dibasic) 500g 71003-498 Tryptone 500g 71003-452 Potassium Phosphate (Dibasic) 1kg 71003-500 Urea, Ultrapure Grade 1kg 95029-278 Potassium Phosphate Dibasic Solution [1M] 1L 82022-850 Urea, Ultrapure Grade 500g 71003-454 Potassium Phosphate (Monobasic) 500g 82023-104 X-Gal (Molecular Biology Grade) 1g 71003-456 Potassium Phosphate (Monobasic) 1kg 71003-502 X-Gal (Molecular Biology Grade) 100mg 95029-280 Potassium Phosphate Monobasic Solution [1M] 1L 95029-328 X-Gluc (Molecular Biology Grade) 100mg 89130-998 RBC Lysis Buffer 100ml 71003-504 Xylene Cyanol 10g 89131-000 RBC Lysis Buffer 250ml 71003-506 Yeast Extract 100g 71003-508 Yeast Extract 500g

28 For further details, visit VWR.com G-Biosciences Product Line Overview

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