AMPLIFICATION PCR Reagents

+ How do you ensure optimal amplification results?

+ How do you choose the best PCR enzyme for your application?

+ How can you improve cDNA synthesis? 0>0 PCR REAGENTS

+ We have the best PCR reagent for your application.

Our expertise in enzyme engineering has produced one of the broadest portfolios of

high-fidelity PCR enzymes available today. This expertise has translated to advances in

high-fidelity reverse transcription PCR. Whether you need the highest accuracy

available, large product yields, or sensitive cDNA synthesis, trust our high-performance

enzymes to answer your amplification challenges.

CONTENTS:

PCR Enzymes & Master Mixes RT-PCR Kits & Reagents 6 Ultra-High Fidelity PCR 13 RT-PCR 7 High Fidelity for TA/UA Cloning 8 High Fidelity for Blunt-End Cloning PCR-Related Kits & Reagents 9 High Sensitivity & High Yield 14 Nucleotides 10 GC-Rich/Complex Targets 14 PCR Purification Kits 11 Long PCR 14 PCR Cloning Kits 12 Routine PCR 15 Ordering Information PCR ENZYMES

Ultra-High Fidelity High Fidelity High Fidelity High Fidelity Format (Cloning & (Contamination (TA/UA Cloning) (Blunt Cloning) Mutagenesis) Control)

™ ® ® PfuUltraa™ Easy-A PfuTurboo PfuTurboo Cx Hotstart Enzyme Hotstart DNA High Fidelity PCR Hotstart DNA Hotstart DNA oning Enzym Polymerase Polymerase

PfuUltraa™ PfuTurbo® Standard Enzyme High-Fidelity DNA Polymerase DNA Polymerase

PfuUltraa™ Easy-A™ PfuTurboo® Master Mix Hotstart PCR High Fidelity PCR Hotstart PCR Master Mix Master Mix Master Mix

GC-Rich/ High Sensitivity/ Format Complex Long PCR Routine PCR High Yield Templates

PicoMaxx™ Herculase® Hotstart Enzyme High Fidelity Hotstart DNA PCR System Polymerase

Herculase® EXL™ Taq2000 ™ Standard Enzyme nhanced DN DNA Polymerase DNA Polymerase Polymerase

PicoMaxx™ Herculase® Master Mix High Fidelity Hotstart PCR PCR Master Mix Master Mix

RT-PCR KITS & REAGENTS

Format High Yield/ High Sensitivity Fidelity

StrataScript™ cDNA Synthesis First Strand Synthesis Kit

StrataScript™ One-Tube RT-PCR One-Tube RT-PCR Kit with Easy-A™ PCR Cloning Enzyme

StrataScript™ Two-Tube RT-P Two-Tube RT-PCR Kit with PicoMaxx™ PCR System PfuUltra™ DNA Polymerase

www.stratagene.com 4>5 PCR REAGENTS

The Importance of High Fidelity

High-fidelity PCR enzymes are valuable for minimizing the introduction of amplification errors in products that will be cloned, sequenced, and expressed. Significant time and effort can be saved by employing high-fidelity amplification procedures that eliminate the need for downstream error-correction steps and minimize the number of clones that must be sequenced in order to obtain error-free constructs or accurate consensus sequences. Moreover, the use of high-fidelity amplification conditions is essential when analyzing small amounts of template DNA or rare molecules in heterogeneous populations. Amplifications employing small amounts of template DNA are especially prone to high mutant frequencies due to PCR-generated errors in early cycles1, 2.

The ArchaeMaxx® Factor Advantage

Because proofreading PCR enzymes can be finicky, we have improved high-fidelity PCR to

be more reliable. Our ArchaeMaxx® Polymerase-Enhancing Factor, found exclusively in our high-fidelity DNA , improves overall PCR performance by eliminating the inhibition of proofreading enzymes caused by incorporation of dUTP, which results from deamination of dCTP during PCR3. Our PCR enzymes formulated with the ArchaeMaxx factor produce higher yields and exhibit greater target length capability.

Superior Reverse Transcription

Our expertise in enzyme engineering also extends to reverse transcriptases. Both MMLV and AMV reverse transcriptases possess substantial RNase H activity that degrades RNA molecules and limits full-length cDNA synthesis and yield. For this reason, RTs that lack RNase H activity are far superior to MMLV and other RNAse H+ RTs. Our StrataScript™ contains a point mutation that effectively abolishes RNase H activi- ty, allowing more efficient production of full-length cDNA than RNase H+ RTs. Comparison of Our PCR Enzymes

PCR EnzymeAccuracy Target Blunt or ArchaeMaxx® Hot Start Master Mix vs. Taq Length 3'-A Ends Advantage Available Available

0-17 kb (genomic) PfuUltra™ High Fidelity DNA Polymerase 18x Blunt +++ 0-15 kb (vector)

0-19 kb (genomic) PfuTurbo® DNA Polymerase 6x Blunt +++ 0-15 kb (vector)

® PfuTurbo Cx DNA Polymerase 6x 0-10 kb Blunt +

Pfu DNA Polymerase, cloned or native 6x 0-5 kb Blunt

Easy-A™ High Fidelity PCR Cloning Enzyme 6x 0-5 kb 3'-A +++

Herculase® DNA Polymerase 3x 0-37 kb Mixed +++

20-37 kb (genomic) EXL™ DNA Polymerase 3x Mixed + 20-50 kb (vector)

PicoMaxx™ High Fidelity PCR System 2x 0-10 kb Mixed +++

SureStart® Taq DNA Polymerase 1x 0-5 kb 3'-A +

Taq2000 ™ DNA Polymerase 1x 0-4 kb 3'-A

Comparison of Our RT-PCR Kits & Reagents

RT-PCR Reagent Target RNase H cDNA One-Tube Two-Tube Length Deficient Synthesis RT-PCR RT-PCR

StrataScript™ First Strand Synthesis Kit 0-9 kb ++

StrataScript™ First-Strand Synthesis Kit 0-9 kb ++ + plus PicoMaxx™ High Fidelity PCR System

StrataScript™ One-Tube RT-PCR Kit 0-5 kb ++ with Easy-A™ PCR Cloning Enzyme

StrataScript™ Reverse Transcriptase 0-9 kb ++

StrataScript™ Two-Tube Kit 0-9 kb ++ + with PfuUltra™ DNA Polymerase

www.stratagene.com 6>7 PCR REAGENTS > PCR ENZYMES & MASTER MIXES Ultra-High Fidelity PCR

The Most Accurate PCR Enzyme Our continuing efforts to improve the fidelity of PCR have resulted in PfuUltra™ High-Fidelity DNA Polymerase, the most accurate PCR enzyme commercially available. PfuUltra DNA poly- merase is ideal for PCR cloning, site-directed mutagenesis, and anywhere sequence accuracy is critical for downstream applications.

Enhanced Performance In addition to ultra-high fidelity, PfuUltra high-fidelity DNA PCR Enzyme Accuracy Target Blunt or ArchaeMaxx® Hot Start Master Mix vs. Taq Length 3'-A Ends Advantage Available Available polymerase exhibits superior PCR performance compared to PfuUltra™ 0-17 kb (genomic) most PCR proofreading enzymes. The addition of our ArchaeMaxx High Fidelity 18x Blunt +++ 0-15 kb (vector) DNA Polymerase polymerase-enhancing factor promotes higher yields, shorter extension times, and greater target length capability (Figure 2).

High Specificity Hotstart Version Highest Fidelity Available The PfuUltra™ Hotstart DNA Polymeraseø,‡ is an antibody-mediated ™ ø,‡ The PfuUltra high-fidelity DNA polymerase contains a hotstart formulation of PfuUltra high fidelity DNA polymerase ***,‡ genetically engineered mutant of Pfu DNA polymerase that that provides higher specificity and reduced background. delivers 300% greater accuracy. A validated and referenced

4 fidelity assay demonstrates that PfuUltra high-fidelity DNA poly- Convenient Master Mix Formulation merase exhibits an average error rate three-fold lower than Pfu For ultra-high fidelity with added convenience and increased DNA polymerase and 18-fold lower than Taq DNA polymerase, throughput, choose PfuUltra™ Hotstart PCR Master Mix ø,‡. making it the most accurate PCR enzyme available (Figure 1). This 2x formulation contains PfuUltra hotstart DNA polymerase, buffer, magnesium, and dNTPs all in one tube for faster setup and more consistent results.

DNA Polymerases ACCURACY x 105 M 0.97 1.5 3.5 6 9 12 17 kb

0 5 10 15 20 25

PfuUltra™ DNA Polymerase

™ ® PfuTurbo and PfuTurbo Cx DNA Polymerases Tgo DNA Polymerase Herculase® DNA Polymerase Pfx/KOD DNA Polymerase Expand™ High Fidelity Advantage-HF™ Polymerase Taq DNA Polymerase

Figure 1 Figure 2 THE MOST ACCURATE PCR ENZYME AVAILABLE AMPLIFIES A WIDE RANGE OF TARGETS PfuUltra™ High-Fidelity DNA Polymerase provides the greatest accuracy of any commercially PfuUltra™ High-Fidelity DNA Polymerase amplifies genomic available enzyme1. Fidelity was measured using our validated and referenced fidelity assay4. DNA targets up to 17 kb. Accuracy = 1/Error Rate. High Fidelity for TA/UA Cloning

Unique Proofreading Formulation Adds A-Overhangs TA or UA cloning requires PCR products with 3'-A overhangs. We created the Easy-A™ High-Fidelity PCR Cloning Enzyme to automatically add A-overhangs for quick and easy high-fidelity cloning into TA/UA vectors.

Combines Cloning Efficiency with High Accuracy The Easy-A™ high-fidelity PCR cloning enzyme ,‡,† is the only PCR Enzyme Accuracy Target Blunt or ArchaeMaxx® Hot Start Master Mix proofreading DNA polymerase formulation to deliver high- vs. Taq Length 3'-A Ends Advantage Format Available throughput TA or UA cloning. PCR products amplified with the Easy-A™ High Fidelity PCR 6x0-5 kb 3'-A +++ Easy-A PCR cloning enzyme can be cloned directly to T- or Cloning Enzyme U-modified vectors with high cloning efficiency and throughput (Figure 3). But unlike Taq DNA polymerase, the Easy-A PCR cloning enzyme exhibits proofreading activity that provides the accuracy level of Pfu DNA polymerase. High Specificity Hotstart Formulation The Easy-A high fidelity PCR cloning enzyme is provided as Fidelity Without Extra Work an antibody-mediated hotstart formulation, providing higher Adapting blunt-ended fragments amplified with proofreading specificity and reduced background. enzymes requires post-PCR addition of 3'-A overhangs prior to the cloning step. This additional step includes a secondary Convenient Master Mix Formulation ™ incubation with Taq DNA polymerase and requires the opening For the highest throughput in TA or UA cloning, choose Easy-A ,‡,† and closing of tubes, which exposes the lab to contamination High-Fidelity PCR Master Mix . This 2x formulation contains the risk. The Easy-A high-fidelity PCR cloning enzyme does not Easy-A PCR cloning enzyme, buffer, magnesium, and dNTPs all require any post-PCR A-addition steps. in one tube for faster setup and more consistent results.

5' 3' 5' 3' Easy-A-Amplified™ A Easy-A-Amplified™ A A PCR Products A PCR Products 3' 5' 3' 5' Add 0.5-1.5 µl of PCR product Add 0.5-1.5 µl of PCR product to T-modified vector to U-modified vector 3' 5' 3' 5' T-vector T T-vector U-vector U U-vector T U 5' 3' 5' 3' Topoisomerase-mediated ligation or conventional ligation Ligation

T PCR A U PCR A Product Product A T T-vector A U U-vector T-vector Ligation complete U-vector Ligation complete

Figure 3 HIGH CLONING EFFICIENCY WITH EASY-A™ PCR CLONING ENZYME Targets amplified with the Easy-A™ High-Fidelity PCR Cloning Enzyme contain 3'-A overhangs, allowing quick and easy cloning into any T- or U-modified vector.

www.stratagene.com 8>9 PCR REAGENTS > PCR ENZYMES & MASTER MIXES High Fidelity for Blunt-End Cloning

Accuracy with Enhanced Performance We invented the high-fidelity PCR market over a decade ago with the original Pfu DNA Polymerase. Since that time, our expertise has led to improvements in high-fidelity performance with PfuTurbo® ® DNA Polymerase and, more recently, PfuTurbo Cx Hotstart DNA Polymerase. These improvements have made high-fidelity PCR more reliable and versatile.

High Specificity Hotstart Version The PfuTurbo® hotstart DNA polymerase ø,‡ is an antibody- PCR Enzyme Accuracy Target Blunt or ArchaeMaxx® Hot Start Master Mix vs. Taq Length 3'-A Ends Advantage Available Available mediated hotstart formulation of PfuTurbo DNA polymerase that

PfuTurbo ® 0-19 kb (genomic) provides higher specificity, reduced background, and enhanced 6x Blunt +++ 0-15 kb (vector) DNA Polymerase yield of challenging systems.

PfuTurbo ® C x 6x0-10 kb Blunt + DNA Polymerase New Mutant Overcomes Uracil Sensitivity Pfu DNA Heat can cause deamination of cytosine to uracil in a DNA Polymerase, 6x0-5 kb Blunt cloned or native strand. To prevent replication of such a mutation, archaeal (proofreading) polymerases typically stop replication just before reaching a uracil residue in the template5. In PCR, this poisoning phenomenon can result in decreased or no product yield.

® **,‡ Robust High-Fidelity Amplification We formulated PfuTurbo Cx hotstart DNA polymerase with The PfuTurbo® DNA polymerase ø,‡ is a special formulation of a novel mutant of Pfu DNA polymerase that can read through cloned Pfu DNA polymerase and our ArchaeMaxx polymerase- a uracil located in the template strand without stalling. Thus

enhancing factor that produces increased PCR product yields PfuTurbo Cx hotstart DNA polymerase improves the overall without affecting fidelity. The enhanced performance of PfuTurbo reliability of high-fidelity PCR and exhibits less finicky, more DNA polymerase allows the use of shorter extension times, robust performance (Figure 4). longer targets, and lower concentrations of DNA template than are suitable for Pfu DNA polymerase. Prevent Carryover Contamination

Because PfuTurbo Cx hotstart DNA polymerase can replicate uracil-containing DNA, it can be used to prevent carry-over contamination. This method involves the use of dUTP in place of dTTP in the nucleotide pool. Subsequent PCR reactions are ® ® ® MPFUTURBO Cx ACCUPRIME PFX PLATINUM PFX pre-treated with Uracil-N-glycosylase (UNG) to degrade any dU-containing DNA left over from a previous reaction; the UNG 9 kb

6 kb is then inactivated during the next denaturation step. PfuTurbo Cx hotstart DNA polymerase incorporates dUTP in targets up to 6 kb 2.6 kb in length more efficiently than Taq DNA polymerase.

0.9 kb

Figure 4 ® SUPERIOR PERFORMANCE WITH PFUTURBO CX HOTSTART DNA POLYMERASE ® Amplification of a range of targets from 0.9 to 9 kb using PfuTurbo Cx Hotstart DNA Polymerase and other commercially available proofreading PCR enzymes. High Sensitivity & High Yield

Greater Performance at a Fraction of the Cost Blending Taq DNA polymerase with a proofreading enzyme improves target-length capability, fidelity, and yield, but many PCR enzyme blends lack sensitivity and can lead to PCR failures. We designed the PicoMaxx™ High Fidelity PCR System to deliver the highest sensitivity and efficiency, bringing you greater blend performance at a much lower cost per unit than other enzyme blends.

Sensitivity and Reliability The PicoMaxx™ high fidelity PCR system ø,‡ is designed to provide PCR Enzyme Accuracy Target Blunt or ArchaeMaxx® Hot Start Master Mix maximum PCR sensitivity and reliability. Sensitivity is critical for vs. Taq Length 3'-A Ends Advantage Format Available successful amplification, especially with limited or precious sam- PicoMaxx™ High Fidelity 2x0-10 kb Mixed +++ ples; use of a PCR enzyme that lacks sensitivity often results in PCR System amplification failures. Formulated with a blend of Taq and Pfu DNA polymerases and our ArchaeMaxx polymerase-enhancing factor, together with a specially optimized buffer, the PicoMaxx high fidelity PCR system overcomes such PCR failures with The ArchaeMaxx factor maximizes the performance of the proof- greater sensitivity than other PCR enzymes (Figure 5). Superior reading component of the blend, allowing the PicoMaxx high sensitivity makes the PicoMaxx high fidelity PCR system one of fidelity PCR system to deliver higher product yields and more the most reliable PCR enzyme formulations available. reliable performance across a wide range of targets up to 10 kb.

High Efficiency and Yield Convenient Master Mix Formulation The PicoMaxx high fidelity PCR system exhibits superior amplifi- For maximum sensitivity and yield with added convenience, ™ ø,‡ cation efficiencies due in large part to our patented ArchaeMaxx choose PicoMaxx High Fidelity PCR Master Mix . This 2x factor, which eliminates dUTP that accumulates during the high formulation contains the PicoMaxx high fidelity enzyme blend, heat of PCR and can poison high-fidelity reactions. buffer, magnesium, and dNTPs all in one tube for faster setup and more consistent results.

PICOMAXX™ PLATINUM® TAQ EXPAND™ HIGH FIDELITY HIGH FIDELITY HIGH FIDELITY Genomic DNA Template Qty. 50 25 10 1 50 25 10 1 50 25 10 1 (ng)

3.9 kb

Figure 5 PICOMAXX™ HIGH FIDELITY PCR SYSTEM EXHIBITS SUPERIOR SENSITIVITY We amplified a 3.9 kb α-1 anti-trypsin template with PicoMaxx™ High Fidelity PCR System, Expand™ High Fidelity PCR System (Roche), and Platinum® Taq DNA Polymerase High Fidelity (Invitrogen). We performed all reactions according to the manufacturer’s recommendations.

www.stratagene.com 10>11 PCR REAGENTS > PCR ENZYMES & MASTER MIXES GC-Rich/Complex Targets

Overcome Challenging PCR Some PCR targets are problematic to amplify due to their content or structure. Our Herculase® Enhanced DNA Polymerase helps overcome these PCR challenges with successful amplification of complex and GC-rich templates over a wide range of template sizes.

High Specificity Hotstart Version The Herculase® Hotstart DNA Polymerase ø,‡ is an antibody- PCR Enzyme Accuracy Target Blunt or ArchaeMaxx® Hot Start Master Mix vs. Taq Length 3'-A Ends Advantage Available Available mediated hotstart formulation of Herculase enhanced DNA

Herculase® polymerase that provides higher specificity, reduced back- 3x0-37 kb Mixed +++ DNA Polymerase ground, and enhanced yield of challenging systems.

Convenient Master Mix Formulation For added convenience and increased throughput, choose GC-Rich and Complex Templates Herculase® Hotstart PCR Master Mix ø,‡. This 2x formulation ® ø,‡ The Herculase enhanced DNA polymerase meets today’s contains Herculase hotstart DNA polymerase, buffer, complex PCR challenges. When you need to accurately amplify magnesium, and dNTPs all in one tube for faster setup and a complex or GC-rich template, Herculase DNA polymerase will more consistent results. help you succeed. A unique optimized formulation of high fidelity Pfu DNA polymerase, Taq DNA polymerase and our ArchaeMaxx polymerase-enhancing factor, Herculase DNA polymerase excels in amplifying a broad range of target lengths with superior fidelity than Taq DNA polymerase and other PCR enzyme blends6. DMSO is provided separately as a PCR adjunct, and can be added when amplifying difficult targets to increase product yield and extend target-length capability (Figure 6).

HERCULASE® COMPETITOR’S GC-RICH PCR ENZYME +/- DMSO (%) +/- GC-Rich Solution (M) 0 6 7 8 90 0.5 1.0 1.5 2.0

M 1 2 3 4 5 6 7 8910

Figure 6 HERCULASE® ENHANCED DNA POLYMERASE EXCELS IN AMPLIFYING GC-RICH TARGETS Herculase® Enhanced DNA Polymerase easily amplifies an 82.5% GC-rich fragment of the Fragile X gene from human genomic DNA. Long PCR

Go the Full Distance To amplify very long targets, a blend of Taq DNA polymerase and a proofreader provides the best results. But not every enzyme blend can go the distance. Our EXL™ DNA Polymerase is specially optimized to successfully and accurately amplify long targets up to 50 kb.

Tackle Long Amplicons The EXL™ DNA polymerase ø,‡ provides superior performance PCR Enzyme Accuracy Target Blunt or ArchaeMaxx® Hot Start Master Mix in amplifying complex targets greater than 20 kb in length. vs. Taq Length 3'-A Ends Advantage Available Available

The EXL DNA polymerase is a special formulation of Pfu DNA EXL™ DNA 20-37 kb (genomic) 3x Mixed + Polymerase 20-50 kb (vector) polymerase, Taq DNA polymerase, and our ArchaeMaxx polymerase enhancing factor, along with a special buffer optimized specifically for very long targets. This enzyme blend provides robust yields and reliable amplification (Figure 7).

Amplify with Higher Accuracy Because PCR enzymes have an intrinsic rate at which they incorporate incorrect bases, long templates are particularly prone to errors when amplified during PCR. Most “Long & Accurate” PCR enzyme blends available commercially provide only a slight increase in accuracy over Taq. EXL DNA poly- merase provides twice the accuracy of these other blends to ensure minimal errors in your long PCR products.

EXL™ COMPETITOR

M2330 45 23 30 45

Figure 7 EXL™ DNA POLYMERASE EXCELS AT AMPLIFYING EXTREMELY LONG TARGETS EXL™ DNA Polymerase easily amplifies two extremely long genomic targets (23 and 30 kb) and a lambda target (45 kb). A competitor’s PCR enzyme for long targets is shown for comparison. All reactions were performed according to the manufacturer’s recommendations.

www.stratagene.com 12>13 PCR REAGENTS > PCR ENZYMES & MASTER MIXES Routine PCR

High-Quality Taq Enzymes Provide Consistent Results When fidelity is not of concern, our Taq2000™ DNA Polymerase provides consistent amplification of simple targets. For applications requiring higher specificity, including real-time QPCR, choose SureStart® Taq DNA Polymerase for hotstart capability.

Hotstart for Maximum Specificity Our SureStart® Taq DNA polymerase‡ is a hot-start Taq DNA PCR Enzyme Accuracy Target Blunt or ArchaeMaxx® Hot Start Master Mix vs. Taq Length 3'-A Ends Advantage Format Available polymerase that can be incorporated into PCR protocols previously

SureStart® Taq optimized with Taq DNA polymerase, with little modification of 1x0-5 kb 3'-A + DNA Polymerase cycling parameters or reaction conditions. This specially modified

Taq2000 ™ Taq DNA polymerase allows setup of PCR reactions at ambient 1x0-14 kb 3'-A DNA Polymerase room temperature without nonspecific annealing and extension during PCR setup, making setup easier. SureStart Taq DNA polymerase can be used in a variety of amplification systems Consistent Performance to improve specificity, yield, and amplification of difficult targets. The Taq2000 ™ DNA polymerase‡ is a highly purified, recombinant Taq DNA polymerase cloned from the thermophilic eubacterium, Efficient, Sensitive, Real-Time PCR Quantification Thermus aquaticus. Using Taq2000 DNA polymerase in longer As part of our rigorous quality control testing, every lot of PCR amplifications reduces smearing and virtually eliminates SureStart Taq DNA polymerase is routinely validated for QPCR. unwanted background artifacts. Taq2000 DNA has superior For maximum performance in real-time PCR, choose one of our α thermostability compared to other commercial Taq DNA Brilliant® QPCR and QRT-PCR reagent kits or master mixes‡, , polymerase preparations. each of which is formulated with SureStart Taq DNA polymerase. RT-PCR REAGENTS > RT-PCR KITS & REAGENTS RT-PCR

Produce High cDNA Yields Most common reverse transcriptases, including MMLV and AMV, exhibit RNase H activity, which limits yields of full-length cDNA. With StrataScript™ Reverse Transcriptase we have eliminated RNase H activity to provide greater yields of full-length transcripts.

Mutant RT Delivers Higher Yields and Sensitivity The StrataScript™ reverse transcriptase is a Moloney murine RT-PCR Target RNase H cDNA One-Tube Two-Tube leukemia virus reverse transcriptase (MMLV RT) that has Reagent Length Deficient Synthesis RT-PCR RT-PCR

been genetically modified to remove RNase H activity without StrataScript™ First Strand 0-9 kb + + affecting the desired reverse transcriptase function. As a result, Synthesis Kit

StrataScript RT generates longer transcripts and greater StrataScript™ One-Tube RT-PCR Kit 0-5 kb + + performance. We manufacture StrataScript RT to be free from with Easy-A™ PCR Cloning Enzyme impurities, allowing for the highest possible cDNA yield (Figure 8). StrataScript™ Two-Tube Kit with 0-9 kb + + + StrataScript RT also provides superior sensitivity compared to PfuUltra™ DNA Polymerase MMLV and AMV reverse transcriptases in real-time QRT-PCR applications. StrataScript RT is the ideal choice to generate cDNA for a variety of downstream applications including library construction and RT-PCR. One-Tube RT-PCR System for Cloning The StrataScript™ One-Tube RT-PCR System‡ combines high High-Quality First-Strand Synthesis cloning efficiency with high fidelity in a one-tube RT-PCR format. The StrataScript™ First Strand cDNA Synthesis Kit§ employs The kit features our robust StrataScript RT and our Easy-A PCR StrataScript RT to generate cDNA of high quality from total or cloning enzyme, which amplifies with six-fold higher accuracy poly(A) RNA. Subsequent amplification with sequence-specific than Taq DNA polymerase yet adds 3'-A overhangs for efficient primers yields a homogenous population of the specific cDNA TA or UA cloning. molecule of interest. Ultra-High Fidelity RT-PCR The StrataScript™ Two-Tube RT-PCR System‡ is the highest fidelity RT-PCR system available. Our StrataScript RT is included for highly efficient first strand synthesis. The cDNA is then amplified with PfuUltra high fidelity DNA polymerase. The StrataScript two-tube RT-PCR system is ideal for applications where sequence integrity is critical.

High Sensitivity Two-Tube RT-PCR When sensitivity is important, and you want to archive your cDNA, combine our StrataScript first strand cDNA synthesis kit with our PicoMaxx™ high fidelity PCR system. The PicoMaxx 100 U 100 U STRATASCRIPT™ SUPERSCRIPT™ II PCR system provides the most sensitive amplification of any RT RT PCR enzyme or blend available.

Figure 8 SAME ROBUST PERFORMANCE Reactions using 100 U of StrataScript™ Reverse Transcriptase and 100 U of SuperScript™ II Reverse Transcriptase (Invitrogen) generated equivalent performance at 40ºC.

www.stratagene.com 14 PCR-RELATED KITS & REAGENTS PCR-Related Kits & Reagents

We offer a wide range of kits and reagents to complete your PCR and prepare for a variety of downstream applications.

High-Quality PCR-Grade Deoxynucleotide Mix Efficient PCR Cloning Our dNTP mix contains 400 µl of 100 mM dNTP mix (25 mM PCR-Script® Cloning Kits§, allow the efficient cloning of PCR each of dATP, dCTP, dGTP, and dTTP), adequate for 1000-2000 fragments with a high yield and a low rate of false positives. standard primer-extension reactions. This mix produces high- PCR products are incubated with one of the predigested quality reaction products and withstands multiple freeze-thaw PCR-Script cloning vectors, Srf I and T4 DNA ligase. Using cycles without compromising efficiency. the restriction enzyme in the ligation reaction maintains a high steady-state concentration of digested vector DNA and allows the Safe, Easy PCR Cleanup use of nonphosphorylated, unmodified PCR primers. The ligation Our StrataPrep® PCR Purification Kits prepare PCR products efficiency of blunt-ended DNA fragments is increased by the quickly and easily for downstream applications such as simultaneous, opposite reactions of the Srf I restriction enzyme sequencing, microarray analysis, and cloning. Following and T4 DNA ligase on nonrecombinant vector DNA. The PCR- amplification or cDNA labeling, a DNA binding solution is added Script cloning kits include the StrataPrep PCR purification kit directly to the PCR tube and then transferred to a unique and also include polishing reagents to create blunt-ended PCR microspin cup. Once bound, the DNA (cDNA) is washed and products amplified with a non-proofreading enzyme. then eluted from the column, ready for resuspension in your application buffer of choice. For high-throughput applications, our StrataPrep® 96 PCR Purification Kit provides the same easy and efficient cleanup in a convenient 96-well format. ORDERING INFORMATION Ordering Information

PCR Enzymes PCR Master Mixes

Product Name Size Catalog Product Name Size Catalog Ultra-High Fidelity Ultra-High Fidelity PFUULTRA™ HIGH FIDELITY DNA POLYMERASE 100 U 600380 PFUULTRA™ HOTSTART PCR MASTER MIX 100 rxn 600630 500 U 600382 400 rxn 600632 1000 U 600384 High Fidelity, TA or UA Cloning PFUULTRA™ HOTSTART DNA POLYMERASE 100 U 600390 EASY-A™ HIGH FIDELITY PCR MASTER MIX 100 rxn 600640 500 U 600392 400 rxn 600642 1000 U 600394 High Fidelity, Blunt Cloning High Fidelity, TA or UA Cloning PFUTURBO ® HOTSTART PCR MASTER MIX 100 rxn 600600 EASY-A™ HIGH FIDELITY PCR CLONING ENZYME 100 U 600400 400 rxn 600602 500 U 600402 High Sensitivity & Yield 1000 U 600404 PICOMAXX™ HIGH FIDELITY PCR MASTER MIX 100 rxn 600650 High Fidelity, Blunt Cloning GC-Rich/Complex Targets ® PFU DNA POLYMERASE, CLONED 100 U 600153 HERCULASE HOTSTART PCR MASTER MIX 100 rxn 600610 500 U 600154 400 rxn 600612 1000 U 600159 PFU DNA POLYMERASE, NATIVE 100 U 600135 RT-PCR Reagents 500 U 600136 1000 U 600140 RT-PCR PFUTURBO® C HOTSTART DNA POLYMERASE 100 U 600410 X STRATASCRIPT™ FIRST STRAND SYNTHESIS KIT 50 rxn 200420 500 U 600412 STRATASCRIPT™ ONE-TUBE RT-PCR KIT 50 rxn 600168 1000 U 600414 WITH EASY-A™ PCR CLONING ENZYME PFUTURBO® DNA POLYMERASE 100 U 600250 STRATASCRIPT™ REVERSE TRANSCRIPTASE, 50U/µL 10,000U 600085 500 U 600252 STRATASCRIPT™ REVERSE TRANSCRIPTASE, 200U/µL 10,000U 600086 1000 U 600254 STRATASCRIPT™ TWO-TUBE RT-PCR KIT 50 rxn 600170 ® PFUTURBO HOTSTART DNA POLYMERASE 100 U 600320 WITH PFUULTRA™ DNA POLYMERASE 500 U 600322 1000 U 600324 High Sensitivity & Yield PCR-Related Kits & Accessories PICOMAXX™ HIGH FIDELITY PCR SYSTEM 100 U 600420 500 U 600422 Nucleotide Mix 1000 U 600424 DEOXYNUCLEOTIDE MIX, PCR-GRADE (G/A/T/C, 25 mM EACH) 400 µl 200415 GC-Rich/Complex Targets PCR Purification HERCULASE® ENHANCED DNA POLYMERASE 100 U 600260 STRATAPREP® 96 PCR PURIFICATION KIT 2 plates 400775 500 U 600262 10 plates 400776 1000 U 600264 50 plates 400774 HERCULASE® HOTSTART DNA POLYMERASE 100 U 600310 STRATAPREP® PCR PURIFICATION KIT 50 preps 400771 500 U 600312 250 preps 400773 1000 U 600314 PCR Cloning Kits Extra-Long Targets PCR-SCRIPT® AMP CLONING KIT 10 rxn 211188 ™ EXL DNA POLYMERASE 100 U 600340 25 rxn 211190 500 U 600342 50 rxn 211189 1000 U 600344 PCR-SCRIPT® CAM CLONING KIT 25 rxn 211192 Routine PCR SURESTART® TAQ DNA POLYMERASE 100 U 600280 500 U 600282 1000 U 600284 TAQ2000™ DNA POLYMERASE 100 U 600195 500 U 600196 1000 U 600197

LEGAL Expand is a trademark of Roche Diagnostics Corporation. * U.S. Patent Nos. 6,734,293, 6,444,428, 6,379,553, 6,333,165, 6,183,997 and patents pending. Platinum is a registered trademark of Invitrogen Corporation. ** U.S. Patent Nos. 6,734,293, 6,444,428, 6,379,553, 6,333,165, 6,183,997, 6,489,150, 5,948,663, 5,866,395, SuperScript is a trademark of Invitrogen Corporation. 5,556,772, 5,545,552 and patents pending. *** U.S. Patent Nos. 6,489,150, 5,948,663, 5,866,395, 5,545,552 and patents pending. REFERENCES Patents pending 1. Hogrefe, H.H. and M.C. Borns. High Fidelity PCR Enzymes. In C.W. Dieffenbach, G.S. Dveksler (eds.) ø U.S. Patent Nos. 6,734,293, 6,489,150, 6,444,428, 6,379,553, 6,333,165, 6,183,997, 5,948,663, 5,866,395, PCR Primer: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 2003. 5,556,772, 5,545,552 and patents pending. 2. Cha, R.S. and W.G. Thilly. Specificity, efficiency, and fidelity of PCR. In PCR Primer: A Laboratory Manual ‡ Purchase of these products is accompanied by a license to use them in the Polymerase Chain Reaction (PCR) process (eds, Dieffenbach, C.W. and G.S. Dveksler) Cold Spring Harbor Laboratory Press, 1995. in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by the 3. Hogrefe, et al. (2002). Proc. Nat. Acad. Sci.USA 99: 596-601. up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., an authorized thermal cycler. 4. Cline, J., Braman, J.C. and Hogrefe, H.H. (1996) Nucleic Acids Res. 24: 3546-3551. § Purchase of this PCR-related product does not convey any rights under the PCR patents owned by Roche Molecular Systems. A license to use the PCR process accompanies the purchase of certain reagents from Stratagene when used 5. Fogg, et al. (2002). Nat Struct Biol. 9(12): 922-927. in conjunction with an Authorized Thermal Cycler. 6. Borns, M. and Hogrefe, H. (2000) Strategies. 13: 12. † Use of these products for certain applications may require licenses from third parties in certain countries. Use of the CMV promoter is covered under U.S. Patent Nos. 5,168,062 and 5,385,839 owned by the University of Iowa Research Foundation and licensed FOR RESEARCH USE ONLY. α Use of labeling reagents may require licenses from entities other than Stratagene. For example, use of fluorogenic probes in 5' nuclease assays may require licenses under U.S. Patent Nos. 6,214,979, 5,804,375, 5,210,015 and 5,487,972 owned by Roche Molecular Systems, Inc. and under U.S. Patent No. 5,538,848 owned by Applied Biosystems.

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