AmpliTaq Gold

Chemically modified hot-start DNA polymerases for enhanced specificity

Applied Biosystems AmpliTaq Gold and AmpliTaq Gold 360 DNA Polymerases are hot-start enzymes which utilize a chemical modification to inhibit enzyme activity at room temperature. Upon activation the modifier is permanently released, resulting in an enzyme that becomes active at a temperature above the primer annealing temperature. Thus the only products extended and amplified are those with highly specific priming.

What is hot-start PCR?

Learn about common issues in PCR amplification and how you can resolve them with hot-start PCR. Also discover different types of hot-start enzyme modifications and how to choose a hot-start DNA polymerase suitable for your PCR.

AmpliTaq Gold product formats

While both AmpliTaq Gold formats are designed for hot-start reactions, the AmpliTaq Gold 360 DNA Polymerase is more versatile because it can better handle GC-rich templates and is more flexible for shipping, storage, and handling. Their comparison to our latest hot-start enzyme, Platinum II Taq Hot-Start DNA Polymerase, is also provided here as it may be better suited for your PCR applications.

  AmpliTaq Gold 360 DNA Polymerase AmpliTaq Gold DNA Polymerase Platinum II Taq Hot-Start DNA Polymerase
Hot-start modification Chemical Chemical Antibody
Enzyme activation time 10 min 10 min 2 min
Universal annealing temperature of 60°C No No Yes
DNA synthesis speed 60 sec/kb 60 sec/kb 15 sec/kb
Amplification length Up to 5 kb Up to 5 kb Up to 5 kb
Inhibitor tolerance No No Yes
GC-rich DNA amplification ++ + +++
Benchtop stability of assembled reactions Yes Yes Up to 24 hr
Certified low level of residual DNA
  • Human gDNA:  ≤2 copies per 10 U
  • E. coli gDNA: ≤1.4 copies per 5 U
 
  • Human gDNA:  ≤0.2 copy per 50 μL reaction
  • Bacterial gDNA: ≤1 copy per 50 μL reaction  
Stand-alone enzyme
Master Mix format Colorless   Colorless
Green*
Storage temperature –20°C** –20°C  –20°C**
Ambient shipping Yes No  No

* Contains electrophoresis tracking dyes and a density reagent for direct gel loading of PCR products.
** Master mixes can be stored at +4°C for up to 3 months after thawing.


What is the difference between AmpliTaq Gold and AmpliTaq Gold 360 DNA Polymerases?

Compared to the original AmpliTaq Gold DNA Polymerase, AmpliTaq Gold 360 DNA Polymerase is purified by an additional proprietary separation process which reduces residual bacterial DNA sequences from the enzyme preparation. When used with the enhanced 360 Gold Buffer, this ultra-pure enzyme, in addition to its hot-start capabilities, reduces false positive results, amplifies low-level target sequences, and promotes the amplification of a variety of templates including those from bacterial and human genomes.

Due to its purity, AmpliTaq Gold 360 DNA Polymerase efficiently amplifies low-copy targets, making it suitable for detection of sequences that are rare or degraded. AmpliTaq Gold 360 DNA Polymerase is able to amplify sequences up to 5 kb.


AmpliTaq Gold 360 DNA Polymerase advantages

Versatility

Figure 1. AmpliTaq Gold 360 DNA Polymerase amplifies a broad range of targets. The panel shows products amplified with AmpliTaq Gold 360 DNA Polymerase. PCR was performed in duplicate with 1 ng of template DNA per reaction. Amplicons ranged from 300 to 1400 base pairs (bp) in length with an average length of 553 bp. Amplicons are labeled as follows: High AT; Easy Amplification; Primer Dimer; Homopolymer; Sequencing Challenge Long; High GC. The high GC amplicon reactions included 5 µL of 360 GC Enhancer.

Sensitivity

Figure 2. Sensitivity of AmpliTaq Gold 360 DNA Polymerase for detection of low copy targets. A gel image showing the amplification products of the ß-actin gene performed with 0 to 80 copies of input DNA, using 2.0 mM MgCl2 and standard thermal cycling conditions.

Quality

Figure 3. High-quality sequencing data obtained from AmpliTaq Gold 360 DNA Polymerase-generated PCR products. Amplification products from both standard (Panel A) and high GC content (Panel B) targets generated by AmpliTaq Gold 360 DNA Polymerase and AmpliTaq Gold DNA Polymerase were sequenced (10 ng PCR product per sequencing reaction). Samples were cycle-sequenced using BigDye Terminator v3.1 and standard thermal cycling conditions. BigDye XTerminator Purification Kit was used to clean up the cycle sequencing reaction, and the samples were injected into the Applied Biosystems 3730xl DNA Analyzer with POP-7 polymer.

 

Versatility

Figure 1. AmpliTaq Gold 360 DNA Polymerase amplifies a broad range of targets. The panel shows products amplified with AmpliTaq Gold 360 DNA Polymerase. PCR was performed in duplicate with 1 ng of template DNA per reaction. Amplicons ranged from 300 to 1400 base pairs (bp) in length with an average length of 553 bp. Amplicons are labeled as follows: High AT; Easy Amplification; Primer Dimer; Homopolymer; Sequencing Challenge Long; High GC. The high GC amplicon reactions included 5 µL of 360 GC Enhancer.

Sensitivity

Figure 2. Sensitivity of AmpliTaq Gold 360 DNA Polymerase for detection of low copy targets. A gel image showing the amplification products of the ß-actin gene performed with 0 to 80 copies of input DNA, using 2.0 mM MgCl2 and standard thermal cycling conditions.

Quality

Figure 3. High-quality sequencing data obtained from AmpliTaq Gold 360 DNA Polymerase-generated PCR products. Amplification products from both standard (Panel A) and high GC content (Panel B) targets generated by AmpliTaq Gold 360 DNA Polymerase and AmpliTaq Gold DNA Polymerase were sequenced (10 ng PCR product per sequencing reaction). Samples were cycle-sequenced using BigDye Terminator v3.1 and standard thermal cycling conditions. BigDye XTerminator Purification Kit was used to clean up the cycle sequencing reaction, and the samples were injected into the Applied Biosystems 3730xl DNA Analyzer with POP-7 polymer.

 

Ordering information

 

Resources

PCR education

Learn about the history of PCR, setup considerations, importance of DNA polymerase choice, common methods and applications, and troubleshooting.

Molecular biology webinars

View our webinar series to learn more about molecular biology topics such as reverse transcription, PCR and cloning.

Support

PCR and cDNA synthesis support center
Find tips, troubleshooting help, and resources for your end-point PCR and cDNA applications.

OEM & customized PCR solutions
Discover customizable manufacturing solutions, product labeling, and packaging capabilities for your specific requirements.

Contact us
Email or call our Technical Application Scientists for additional questions regarding PCR enzymes and master mixes.