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AmpliTaq Gold™ DNA Polymerase with Gold Buffer and MgCl2 (Applied Biosystems™)

AmpliTaq Gold® DNA Polymerase is a chemically modified form of AmpliTaq® DNA Polymerase requiring thermal activation. Features of this enzyme:

• Automated chemical hot-start enzyme for increased specificity, sensitivity, and yield
• Time-released thermal activation improves sensitivity in low copy number amplifications
• Successful multiplex PCR saves time and reagents
• Includes GeneAmp® 10X PCR Gold Buffer with MgCl2

Hot-start activation
The modified enzyme is provided in an inactive state. Upon thermal activation, the modifier is permanently released, regenerating active enzyme. The resulting hot-start PCR amplification provides increased sensitivity, specificity, and yield over conventional PCR techniques.

Higher specificity, higher yield
AmpliTaq Gold® DNA Polymerase can be activated partially or completely in a pre-PCR heat step, or can be allowed to activate slowly in a time-released manner during the denaturation steps of thermal cycling. With or without a limited up-front heat activation step, active enzyme is released slowly during thermal cycling to match template concentration and increase specificity. The yield of specific product increases because reactants are not wasted in the formation of unintended products. Because AmpliTaq Gold® DNA Polymerase is a chemical hot-start enzyme, there is no worry of biological contamination.

GeneAmp® 10X PCR Gold Buffer is formulated to provide flexible, efficient activation of AmpliTaq Gold® DNA Polymerase, resulting in a highly specific, robust PCR amplification. The ionic strength and the pH of GeneAmp® 10X PCR Gold Buffer have been optimized to provide a wider activation temperature and time range when used in conjunction with AmpliTaq Gold® DNA Polymerase.

AmpliTaq Gold® 360 DNA Polymerase for superior performance
Also available is AmpliTaq Gold® 360 DNA Polymerase for even better PCR performance. Compared to AmpliTaq Gold® DNA Polymerase, AmpliTaq Gold® 360 DNA Polymerase is purified by an additional proprietary separation process to eliminate 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. Also included with this product is a 360 GC Enhancer to work through challenging GC-rich templates.

AmpliTaq Gold™ 360 Master Mix (Applied Biosystems™)

AmpliTaq Gold® 360 PCR Master Mix contains everything required for successful PCR amplification in one convenient package—all components are premixed and premeasured. AmpliTaq Gold® 360 Master Mix was designed for 360° coverage of a full range of targets. The master mix is supplied at 2X the recommended usage concentration for easy dilution when adding template and primers. Designed for convenience, the AmpliTaq Gold® 360 PCR Master Mix scales to various reaction volumes for greater application and format flexibility. The main ingredient of the master mix is AmpliTaq Gold® 360 DNA Polymerase but it also contains the world-class 360 GC Enhancer which can be added optionally for high GC–content templates. With the wide template coverage, optimization of PCR reaction conditions, is virtually eliminated with the use of AmpliTaq Gold® 360 Master Mix. Key features:

• 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
• Easy-to-use, premixed master mix

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 (see figure). 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 (see table).

Optimized for automated, hot-start PCR
AmpliTaq Gold® 360 DNA Polymerase is the key ingredient in an automated, convenient, and efficient hot-start PCR. When AmpliTaq Gold® 360 PCR Master Mix is added to the reaction mixture at room temperature, the inactive enzyme is not capable of primer extension. Any low-stringency mispriming events that may have occurred will not be enzymatically extended and subsequently amplified. An initial thermal incubation step is required for activation and ensures that active enzyme is generated only at temperatures where the DNA is fully denatured. 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 (see figure). The extreme specificity allows easier multiplexing and allelic discrimination.

Amplify low-copy amplicons and long targets
AmpliTaq Gold® 360 DNA Polymerase efficiently amplifies targets present at low copy number (see figure), 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. The figure shown demonstrates robust PCR amplification of long human and plasmid DNA.

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