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 Polymerase 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 polymerases, 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™ Reverse Transcriptase 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