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Molecular Biology Essentials molecular biology essentials TaqMan® is a registered trademark of Roche Molecular Systems, Inc. Used under permission and license. you may know us better than you think Look closer—your favorite life sciences products are now by Life Technologies For years, you’ve trusted our products and services for your everyday life sciences needs. As Life Technologies, we’re delivering that same quality in a way that’s simple. Today, one partner for scientifi c innovation. Soon, one source for ordering, service, and support. We’re one company driven to help you advance discovery. www.lifetechnologies.com/one ©2011 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners, unless otherwise noted. TaqMan® is a registered trademark of Roche Molecular Systems, Inc. Used under permission and license. real-time PCR 1 nucleic acid purification and analysis 2 end-point PCR 3 cloning 4 RNAi, epigenetics, and non-coding RNA research 5 protein expression, isolation, and analysis 6 ArcturusXT™ laser capture microdissection system 7 I introduction 2011 Life Technologies Molecular Biology Essentials Catalog Comprising trusted and familiar Invitrogen™ and Applied Biosystems® products, the Life Technologies molecular biology portfolio offers life sciences researchers the most-published, high-performance, scalable solutions across the breadth of molecular biology applications. The chart below shows an organized snapshot of these solutions, which have been cited in more than 200,000 publications within the past 10 years. To help you quickly find products that meet your needs, we’ve included icons to denote benefits like introduction time-savings and high-throughput. Refer to the chapter noted for complete product information. Gene analysis and cloning Protein expression • PureLink® Plasmid Prep (Ch. 2) • Invitrogen™ Vector NTI® Cloning & Sequence • PureLink® RNA Mini Kit (Ch. 2) Analysis Software (Ch. 4) Sample • BenchPro® 2100 Plasmid Purification • BenchPro® 2100 Plasmid Purification System preparation System (Ch. 2) • MagMax™ sample preparation system (Ch. 2) • iPrep™ Purification Instrument (Ch. 2) • GeneArt® Gene Synthesis and Gene Assembly • Invitrogen™ protein expression vectors (Ch. 4) including Gateway® vectors (Ch. 6) • E. coli, baculoviral, lentiviral, adenoviral, yeast, Invitrogen™ cloning & mammalian expression systems (Ch. 6) Key • Invitrogen™ Lipofectamine® transfection • SuperScript® Reverse Transcriptase, TOPO® technologies reagents (Ch. 6) cloning, Gateway,® TA Cloning® (Ch. 4) • Neon® Transfection System (Ch. 6) • Platinum® Taq, AmpliTaq Gold® 360 Master • MembranePro™ Functional Protein Expression Mix (Ch. 3) Kit (Ch. 6) • E-Gel® DNA & RNA separation gels (Ch. 2) • Invitrogen™ Qubit® DNA & RNA Quantitation • Tali™ Image-Based Cytometer Detection System (Ch. 2) (information at www.invitrogen.com) • Applied Biosystems® Veriti™, 9700, 2720 thermal • Countess® Automated Cell Counter instruments cyclers (Ch. 3) (information at www.invitrogen.com) • TrackIt™ DNA Ladders (Ch. 2) • Invitrogen™ One Shot® Competent Cells (Ch. 4) • Oligos/primers (Ch. 3), dNTPs (Ch. 1), • imMedia™ growth medium & LB broth (Ch. 4) Invitrogen™ One Shot® competent cells (Ch. 4), • Selection antibiotics (Ch. 6) Essentials UltraPure™ agarose & reagents (Ch. 2) • Ambion® RNA Essentials (Ch. 2), ligases & restriction enzymes (Ch. 4) Time saver High throughput For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. © 2011 Life Technologies Corporation. All rights reserved. II The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. introduction You may notice that we’ve included only our most popular products and featured new technologies within these pages. introduction Follow the web link within the chapters for a complete view of all our products. While you’re on our website, be sure to check out our new products, protocols, technical guides, videos, and handbooks at www.invitrogen.com. If you have any questions about which of our solutions is right for you, please contact Life Technologies technical support by phone, email, Facebook, or Twitter at www.facebook.com/LifeTechnologies and twitter.com/LifeTechSupport. Functional genetic analysis Protein analysis • TRIzol® Reagent (Ch. 2) • Dynabeads® streptavidin magnetic beads • Ambion® Cells-to-CT™ (Ch. 2) (Ch. 6) • Ambion® mirVana™ miRNA Isolation Kit (Ch. 2) • Invitrogen™ Qubit® Protein Quantitation • Ambion® mirVana™ PARIS™ Kit (Ch. 2) System (Ch. 6) • ArcturusXT ™ Laser Capture Dissection System (Ch. 7) • IP antibodies (Ch. 5) • TaqMan® and SYBR® master mixes (Ch. 1) • Novex® and NuPAGE® Gels (Ch. 6) • TaqMan® Gene Expression, Genotyping (SNP), and CNV • XCell SureLock® gel box (Ch. 6) Assays (Ch. 1) • TaqMan® Protein Assays (Ch. 1) • TaqMan® Non-coding RNA and Ambion® Pre-miR Assays (Ch. 5) • Epigenetics: methylation, histone/chromatin remodeling reagents (Ch. 5) • Ambion® Silencer® Select, Lipofectamine® RNAiMAX reagents, mirVana™ miRNA Mimics and Inhibitors (Ch. 5) Applied Biosystems® Real-Time PCR Instruments • iBlot® Dry Blotting System (Ch. 6) • StepOnePlus™ System & 7500 Fast System (Ch. 1) • BenchPro® 4100 Western Processing Station • ViiA™ 7 Real-Time PCR System (Ch. 1) (Ch. 6) • OpenArray® System (Ch. 1) • Applied Biosystems® primers (Ch. 1) • Novex® Protein Stains (Ch. 6) • SYBR® Green assays (Ch. 1) • Novex® Buffers and protein standards (Ch. 6) • SuperScript® VILO kits (Ch. 3) • Novex® Gel cassettes and pour-your-own essentials (www.invitrogen.com/novexcassettes) Life Technologies offers products and services across many applications. Visit our website to find out more: Cell Culture solutions, go to www.invitrogen.com/cellculture. Cell and Tissue Analysis solutions, go to www.invitrogen.com/cellularanalysis. Sequencing Solutions, go to www.lifetechnologies.com/sequencing. www.lifetechnologies.com III real-time PCR 1 real-time PCR chapter section chapter Overview of real-time PCR 5 Real-time PCR instruments 8 Real-time PCR instrument selection guide 8 Applied Biosystems® OpenArray® Real-Time PCR Platform 9 1 Applied Biosystems® ViiA™ 7 Real-Time PCR System 11 chapter 1 contents chapter Applied Biosystems® 7500 Fast Real-Time PCR System 12 Applied Biosystems® 7500 Real-Time PCR System 13 StepOnePlus™/StepOne™ Real-Time PCR Systems 14 Applied Biosystems® 7900HT Fast Real-Time PCR System 16 High Resolution Melt (HRM) Software 17 RealTime StatMiner® Software 17 Real-time PCR reagents and kits 18 Master mix selection guide 18 TaqMan® Fast Advanced Master Mix 19 TaqMan® Universal Master Mix II 20 TaqMan® PreAmp Master Mix 21 TaqMan® Sample-to-SNP™ Kit 22 TaqMan® Genotyping Master Mix 23 MeltDoctor™ HRM Master Mix 24 Fast SYBR® Green Master Mix 25 Power SYBR® Green PCR Master Mix 26 Express One-Step SuperScript® qRT-PCR Universal 27 TaqMan® Fast Virus 1-Step Master Mix 28 Power SYBR® Green RNA-to-CT™ 1-Step Kit 29 TaqMan® Exogenous Internal Positive Control Reagents 30 Other control reagents, enzymes, primers, and reagents 31 Spectral calibration kits 33 TaqMan® RNase P Instrument Verification Plates 34 www.lifetechnologies.com 2 real-time PCR Gene expression 35 TaqMan® Gene Expression Assay selection guide 35 TaqMan® Gene Expression Assays 36 Lyophilized TaqMan® Gene Expression Assays with Master Mix 37 1 Custom TaqMan® Gene Expression Assays 38 Custom Plus TaqMan® RNA Assays 38 TaqMan® RNase P Detection Reagents 39 TaqMan® Endogenous Controls 39 TaqMan® Array Gene Signature Plates 41 Custom TaqMan® Array Plates 42 TaqMan® Custom Plating Service 43 chapter 1 contents chapter Custom TaqMan® Array Microfluidic Cards 44 TaqMan® Array Gene Signature Microfluidic Cards 45 TaqMan® OpenArray® Real-Time PCR Plates 46 Custom TaqMan® probes and primers 48 GeneAssist™ Pathway Atlas 49 Custom TaqMan® Assay Design Tool 49 TaqMan® MicroRNA and Non-coding RNA Assay selection guide 50 TaqMan® MicroRNA Assays 51 Custom TaqMan® Small RNA Assays 51 TaqMan® Non-coding RNA Assays 52 TaqMan® Pri-miRNA Assays 52 Megaplex™ Primer Pools 53 TaqMan® MicroRNA Reverse Transcription Kit 53 TaqMan® Array Human and Rodent MicroRNA Cards 55 TaqMan® OpenArray® MicroRNA Panels 56 TaqMan® Protein Assays 57 DataAssist™ Software 58 ProteinAssist™ Software 58 3 real-time PCR chapter section chapter Genetic variation 59 TaqMan® SNP genotyping selection guide 59 TaqMan® SNP Genotyping Assays 60 Custom TaqMan® SNP Genotyping Assays 61 TaqMan® Drug Metabolism Genotyping Assays 61 1 GeneAssist™ Copy Number Assay Workflow Builder 61 chapter 1 contents chapter TaqMan® Copy Number Assays 62 TaqMan® OpenArray® Genotyping Plates 64 TaqMan® OpenArray® Digital PCR Kits 66 TaqMan® Mutation Detection Assays 68 TaqMan® Genotyper™ Software 69 CopyCaller™ Software 69 Custom TaqMan® Assay Design Tool 69 www.lifetechnologies.com 4 real-time PCR Overview of real-time PCR Advantages of real-time PCR Quantitative real-time PCR, also called qPCR, combines accumulates during PCR. A single-stranded DNA probe is PCR amplification and detection into a single step, thus constructed containing a reporter fluorescent dye on the eliminating the need to detect products using gel electro- 5´ end and a quencher dye on the 3´ end. While the probe phoresis and enabling the PCR method to be truly quanti- is intact, the proximity of the quencher dye greatly reduces tative. Real-time PCR systems utilize fluorescent dyes to the fluorescence emitted by the reporter dye by fluores- detect the accumulation of PCR
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