Applied Biosystems AmpliTaq DNA Polymerases are recombinant Taq DNA Polymerases used in standard research applications such as routine PCR, genotyping, and colony PCR. The AmpliTaq DNA Polymerases are available in different formulations—AmpliTaq DNA polymerase, AmpliTaq 360 DNA polymerase, AmpliTaq Gold DNA polymerase, and AmpliTaq Gold 360 DNA Polymerase.

While AmpliTaq 360 and AmpliTaq DNA Polymerase are recombinant Taq polymerases, AmpliTaq Gold and AmpliTaq Gold 360 DNA Polymerases are hot-start polymerases. These hot-start enzymes use a chemical modification to inhibit enzyme activity at room temperature. Upon activation, the modifier is released and the enzyme is activated. The activation temperature is typically above the primer annealing temperature, which helps ensures specific priming.

Watch the video on how Amplitaq Gold DNA Polymerases work.

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. Compared to the original AmpliTaq Gold DNA Polymerase, AmpliTaq Gold 360 DNA Polymerase is purified by an additional process which reduces residual bacterial DNA sequences from the enzyme. When used with the 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.

Table 1. Comparison of our hot start polymerases.

 

 

product package and four tubes of reagents

AmpliTaq Gold 360 DNA Polymerase

product package and three tubes of reagents

AmpliTaq Gold DNA Polymerase

product package and three tubes of reagents

Platinum II Taq Hot-Start DNA Polymerase

Hot-start modificationChemicalChemicalAntibody
Enzyme activation time10 min10 min2 min
Universal annealing temperature of 60°CNoNoYes
DNA synthesis speed60 sec/kb60 sec/kb15 sec/kb
Amplification lengthUp to 5 kbUp to 5 kbUp to 5 kb
Inhibitor toleranceNoNoYes
GC-rich DNA amplification++++++
Benchtop stability of assembled reactionsYesYesUp 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
N/A
  • Human gDNA:  ≤0.2 copy per 50 μL reaction
  • Bacterial gDNA: ≤1 copy per 50 μL reaction  
Stand-alone enzyme
  • With 10X AmpliTaq Gold 360 Buffer, 25mM MgCl2, and GC Enhancer
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  • With 10X PCR Gold Buffer and 25mM MgCl2 
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  • With 10X PCR Buffer I (MgCl2 pre-added)
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  • With 10X Buffer II and 25mM MgCl2
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  • With 5X Platinum II PCR Buffer (MgCl2 pre-added) and Platinum GC Enhancer
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Master mix formatColorless
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Colorless
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Green*
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Storage temperature–20°C**–20°C –20°C**
Ambient shippingYesNo 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.

AmpliTaq Gold 360 DNA Polymerase advantages

AmpliTaq Gold 360 DNA Polymerase amplifies a broad range of targets

Gel image with clear bands/amplified targets

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 1,400 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.

AmpliTaq Gold 360 DNA Polymerase can detect low copy targets

Gel image with clear bands

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

High-quality sequencing data obtained from AmpliTaq Gold 360 DNA Polymerase-generated PCR products

Figure 3. High-quality sequencing data obtained from AmpliTaq Gold 360 DNA Polymerase-generated PCR products. Amplification products from both standard (A) and high GC content (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

AmpliTaq Gold 360 DNA Polymerase amplifies a broad range of targets

Gel image with clear bands/amplified targets

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 1,400 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

AmpliTaq Gold 360 DNA Polymerase can detect low copy targets

Gel image with clear bands

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

Sequencing data quality

High-quality sequencing data obtained from AmpliTaq Gold 360 DNA Polymerase-generated PCR products

Figure 3. High-quality sequencing data obtained from AmpliTaq Gold 360 DNA Polymerase-generated PCR products. Amplification products from both standard (A) and high GC content (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.

 

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