AmpliTaq Gold

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.

AmpliTaq Gold product formats

While both formats are designed for hot-start reactions, the AmpliTaq Gold 360 polymerase is especially versatile because it is able to better handle GC-rich templates and is more flexible for shipping, storage, and handling.

  AmpliTaq Gold 360 DNA Polymerase1 AmpliTaq Gold DNA Polymerase
Hot start modification Chemical Chemical
Amplicon length Up to 5 kb Up to 5 kb
Polymerase activation step 10 minutes at 95°C 10 minutes at 95°C
GC-rich sequence amplification Yes No
Stand-alone enzyme
Storage temperature –20°C or +4°C –20°C 
Ambient shipping Yes No 
Master Mix format  Yes No

1 AmpliTaq Gold 360 DNA Polymerase is the same enzyme as AmpliTaq Gold DNA Polymerase, but is further purified and certified to contain low levels of residual DNA: Human DNA ≤2 copies/10 U and Bacterial DNA ≤1.4 copies E.coli DNA/ 5 U.


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.

 

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