AmpliTaq Gold® 360 DNA Polymerase

Catalog number: 4398894

Applied Biosystems™  Related applications: Methylation Analysis | PCR

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Description

The Applied Biosystems® AmpliTaq Gold® 360 DNA Polymerase, when used with improved AmpliTaq Gold 360 Buffer, and the optional 360 GC Enhancer, amplifies a vast range of DNA sequence contexts. AmpliTaq Gold® 360 DNA Polymerase delivers 360° coverage for a full range of targets.

* Optimized for the broadest range of targets—from everyday to challenging
* Unmatched sensitivity, specificity, and yield
* Robust amplification of GC-rich sequences with market-leading 360 GC Enhancer
* Achieves the highest quality sequencing data

Optimized for Easy and Challenging Targets
Challenging targets include AT-rich, GC-rich, primer-dimer forming amplicons, homopolymer repeats, and amplicons that pose sequencing challenges. Amplicons that previously required specialized enzymes and reaction conditions can now be reproducibly amplified with a single reagent under standardized conditions (Figure 1). Competitive benchmarking across more than 40 amplicons distinguishes AmpliTaq Gold® 360 as the best-performing enzyme, ensuring the highest probability of success for the amplification of both everyday and challenging targets (Table 1).

As shown in Figure 1, GC-rich regions are especially poorly amplified with competitor DNA polymerases and the original AmpliTaq Gold, while AmpliTaq Gold 360 provides successful, robust amplification.

Optimized for Hot-Start PCR
AmpliTaq Gold 360 DNA Polymerase provides the same hot-start specificity as AmpliTaq Gold DNA polymerase. A high-temperature incubation step is required to activate AmpliTaq Gold DNA Polymerase, which ensures that the active enzyme is generated only at temperatures in which the DNA is fully denatured and when the primers are not annealed.
When the polymerase is added to the reaction mixture at room temperature, primer extension does not occur because the enzyme is inactivated. Any low stringency mispriming events that may have occurred will not be enzymatically extended and will not be amplified. Hence, PCR setup can be performed at room temperature without concern for extension at misprimed sites. The amount of AmpliTaq Gold 360 DNA polymerase increases in the reaction slowly with each cycle number, and specific product yield increases without buildup of nonspecific products, including primer dimers. Excellent specificity across a broad range of targets is shown in Figure 1 and is summarized in Figure 2. The exquisite specificity allows easier multiplexing and allelic discrimination.

The amount of AmpliTaq Gold 360 DNA polymerase increases in the reaction slowly with each cycle number, and specific product yield increases without buildup of nonspecific products, including primer dimers. Excellent specificity across a broad range of targets is shown in Figure 1 and is summarized in Figures 2A & B. The exquisite specificity allows easier multiplexing and allelic discrimination.

Superior Sensitivity and Amplification Length
Compared to the original AmpliTaq Gold® DNA Polymerase, AmpliTaq Gold® 360 DNA Polymerase is purified by an additional proprietary separation process which reduces contaminating 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.

AmpliTaq Gold 360 DNA Polymerase efficiently amplifies targets present at low copy number (Figure 3), even in the presence of high concentrations of complex DNA, making it especially suited for low-copy pathogen detection, and amplification of targets from degraded DNA samples. The extreme purity of the enzyme contributes to its unmatched sensitivity. AmpliTaq Gold 360 DNA Polymerase efficiently and reproducibly amplifies long (up to 5 Kb) sequences. Figure 4 demonstrates robust PCR amplification of long human and plasmid DNA.

Note: See user's manual or package insert for limited label license, and trademark information. For Research Use Only. Not for use in diagnostics procedures.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Polymerase: AmpliTaq Gold® 360 DNA Polymerase
Hot Start: Built-In Hot Start
Fidelity (vs. Taq): 1 X
GC-Rich PCR Performance: High
Reaction Format: Separate Components
Product Overhang: 3'-A
Reaction Speed: Standard
Final Product Size(s): 5 kb or less
Product Size: 3,000 units
Exonuclease Activity: 5' - 3'

Contents & storage

AmpliTaq Gold® 360 DNA Polymerase is supplied at 5U⁄µL with 10X AmpliTaq Gold® 360 PCR Buffer, 25mM M2Cl2 , and 360 GC Enhancer. Typically, 1.5mL of each of AmpliTaq Gold 360 Buffer, 25mM MgCl2 , and 360 GC Enhancer is supplied with each 250U of enzyme.
• Storage Buffer Enzyme is supplied in 20 mM Tris, pH 9.0, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% (w⁄v) Tween 20,
50% (v⁄v) glycerol
• Source: Recombinant, modified form of the Thermus aquaticus DNA polymerase gene expressed in E. coli

All solutions are prepared in deionized, filter-sterilized water. When stored at -15 to -25 °C, this product maintains performance at least until the expiration date on the tube label. Before use, thaw frozen components to room temperature and mix well.

Product is guaranteed up to 12 months as indicated by the expiration date.

Documents

Manuals & protocols