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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.



  AmpliTaq® 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
 
AmpliTaq&reg 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 sec
2–4 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 min
97.5ºC/10 min
95ºC/40 min
92.5ºC/>130 min
97.5ºC/10 min
95ºC/40 min
92.5ºC/>130 min
97.5ºC/10 min
95ºC/40 min
92.5ºC/>130 min
97.5ºC/10 min
95ºC/40 min
97.5ºC/2 min
95ºC/20 min
Cycling 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
* Assumption of 95°, 15 seconds denaturation per cycle
** pH optimum measured at 72°C