PCR Reagents Selection Guide
Applied Biosystems offers a complete suite of general PCR and reverse-transcriptase PCR reagents to fit with all of your applications. With your new and improved formulation of the worlds' most trusted brand of DNA polymerase, you know you will get the results you are looking for every time.
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PCR Enzymes
* Assumption of 95°, 15 seconds denaturation per cycleAmpliTaq® 360 DNA Polymerase AmpliTaq Gold® 360 DNA Polymerase AmpliTaq Gold® 360 Master Mix AmpliTaq Gold® Fast PCR Master Mix, UP GeneAmp® High Fidelity PCR System rTth DNA Polymerase, XL 
Applications Routine PCR applications including: cloning, genotyping, sequencing, viral gene amplification Amplifies a broad range of template types for hot start PCR applications including: multiplex PCR, cloning, genotyping, sequencing, viral gene amplification Amplifies a broad range of template types for hot start PCR applications including: multiplex PCR, cloning, genotyping, sequencing, viral gene amplification Hot start fast PCR especially designed for sequencing applications Long templates and increased fidelity for accurate PCR in applications such as; cloning, mutagenesis, sequencing Designed for extra long templates, contains both 5'-3' DNA polymerase and 3'-5' exonuclease (proofreading) activities to help ensure efficient XL PCR amplification Amplicon Size < 3 kb < 3 kb < 3 kb < 1.7 kb < 10 kb < 40 kb Amplify wide target range •••• •••• ••••• •••• ••• •• Specificity •• ••• ••• ••• •••• •••• Sensitivity •• •••• •••• •••• •••• ••• Yield • ••• •••• ••• ••• •••• Convenience • •••• ••••• ••••• ••••• ••••• Fidelity ••• ••• ••• ••• ••••• •• Fast •• • •• •••• • •• GC-Rich Templates •••• •••• ••••• •• •• •• PCR Performance Guarantee Yes Yes Yes Yes Yes Yes PCR Technical Support Yes Yes Yes Yes Yes Yes Recombinant Enzyme Yes Yes Yes Yes Yes Yes Molecular Weight 94 kDa 94 kDa N/A 94 kDa 94/92 kDa N/A Number of Amino Acids 832 832 832 832 832/770 N/A Optimal Extension Temperature 72–80 ºC 72–80 ºC 68–72 ºC 72–80 ºC 72–80 ºC Extension Rate (72ºC) 2–4 kb/min 2–4 kb/min 2–4 kb/min 500 bp/5 sec
1 kb/15 sec 1.5 kb/30 sec2–4 kb/min 2–4 kb/min Hot Start Manual Automated Automated Automated Manual Manual Enzyme Activation N/A 2–10 min at 95ºC or <10 additional cycles 2–10 min 10 min N/A N/A Reverse Transcriptase Activity Minimal/Low Minimal/Low Minimal/Low Minimal/Low Minimal/Low Yes, Mn++ dependent Half Life 97.5ºC/10 min
95ºC/40 min 92.5ºC/>130 min97.5ºC/10 min
95ºC/40 min
92.5ºC/>130 min97.5ºC/10 min
95ºC/40 min
92.5ºC/>130 min97.5ºC/10 min
95ºC/40 min
92.5ºC/>130 min97.5ºC/10 min
95ºC/40 min97.5ºC/2 min
95ºC/20 minCycling Half Life* 160 cycles 160 cycles 160 cycles 160 cycles 160 cycles 80 cycles Magnesium Ion PCR Optimum 1–4 mM 1.5–4 mM N/A N/A 1–5 mM 1.5–2.5 mM pH Optimum for PCR Amplification** 7.0–7.5 7.0–7.5 N/A N/A 7.0–7.5 7.0–7.5 pH Optimum for Activation N/A 8.05 at room temperature N/A N/A N/A N/A dNTP PCR Optimum 40–200 µM 40–200 µM N/A N/A 40–200 µM 40–200 µM KCl PCR Optimum 50 mM 50 mM N/A N/A N/A 75–100 mM Primer PCR Optimum 0.1–1.0 µM 0.1–1.0 µM 0.1–1.0 µM 0.2–1.0 µM 0.3 µM 0.4 µM Processivity 50–60 bases 50–60 bases 50–60 bases 50–60 bases 50–60 bases 30–40 bases Longest PCR Product ~5 kb plasmid DNA/~2 kb gDNA ~5 kb plasmid DNA/~2 kb gDNA ~5 kb plasmid DNA/~2 kb gDNA ~1.7 kb gDNA ~10 kb plasmid DNA/~5 kb gDNA >40 kb Extra A Addition Yes Yes Yes Yes Mixture Mixture 5'- 3' Exonuclease Activity Yes Yes No Yes Yes Yes 3'- 5' Exonuclease Activity No No No No Yes Yes Contaminating Nucleases No No No No No No Supplied Concentration 5 Units/µL 5 Units/µL 2X 2X 5 Units/µL 2 Units/µL Storage Temperatures -20ºC -20ºC 4ºC or -20ºC 4ºC -20ºC -20ºC
** pH optimum measured at 72°C -
RT-PCR Kits
High Capacity RNA-to-cDNA Master Mix High Capacity RNA-to-cDNA Kit GeneAmp® Gold RNA PCR Reagent Kit GeneAmp® EZ rTth RNA PCR Kit GeneAmp® Thermostable rTth Reverse Transcriptase RNA PCR Kit 
Application Single-stranded cDNA synthesis for quantitative real-time PCR Single-stranded cDNA synthesis for quantitative real-time PCR RT-PCR with hot-start PCR RT-PCR screening RT-PCR with gene specific primers Hot Start for Reverse Transcriptase No No No No No Hot Start for PCR No No Yes No No Thermostable/ Thermoactive Reverse Transcriptase No No No Yes Yes Reverse Transcriptase Enzyme MuLV Reverse Transcriptase MultiScribe™ Reverse Transcriptase MultiScribe™ Reverse Transcriptase rTth DNA Polymerase rTth DNA Polymerase PCR Enzyme — — AmpliTaq Gold® DNA Polymerase rTth DNA Polymerase rTth DNA Polymerase dUTP/UNG Compatible for Carry-over Prevention — — No Yes Yes Oligos Used Random and oligo d(T) primers Random octomers, oligo d(T)16 Random hexamers, oligo d(T)16, or gene specific primers Gene specific primers Gene specific primers Fidelity +++ +++ +++ ++ +++ Comments Complete reverse transcription master mix formulation, ready for high throughput real-time RT-PCR Two tube configuration reverse transcription, minimal pipetting for two-step real-time RT-PCR application Greatest flexibility for RT-PCR. Use with random hexamers,oligo d(T)16, or gene specific primers. Allows for a one or two step protocol. Incorporation of contaminant control essential for screening of multiple samples to minimize PCR carry-over Optimal for amplification of RNAs containing a high degree of secondary structure and
GC-rich regions due to the fact that reverse transcription is
performed above