Advantage HD Polymerase Mix

The Advantage HD Polymerase Mix provides maximum fidelity and extended PCR product length, making it particularly useful for applications that require high accuracy, such as cDNA cloning, site-directed mutagenesis, and mutation genotyping (i.e., SNP analysis).

The Advantage HD Polymerase Mix consists of a novel DNA polymerase with a hot-start antibody, and comes with a tube of 5X buffer (with Mg2+).

Advantage HD Polymerase Mix provides outstanding accuracy due to the presence of 3' → 5' exonuclease activity that results in an extremely low error rate. It also performs extremely well on GC-rich and other more complex templates. The Advantage HD Polymerase Mix amplifies targets of up to 8.5 kb from human genomic DNA, 10 kb from E. coli genomic DNA, and 22 kb from lambda DNA. The amplification efficiency of Advantage HD Polymerase Mix is greater than that of wild-type Taq polymerase.

Overview

Extremely low error rate—exonuclease proofreading yields just 12 errors per 250,000 bp, and performs well on GC-rich templates.
Integrated hot-start antibody for enhanced specificity—prevents false initiation during reaction setup and minimizes primer-dimer formation, thus reducing background.
High priming efficiency—shortens reaction time and produces results fast.
Direct cloning—PCR products can be directly cloned into blunt-end vectors. If primers having sufficient vector sequence homology are used, then products can be cloned directly using our In-Fusion Cloning technology.

Applications

In-Fusion Cloning for directional, seamless cloning of any PCR fragment or multiple fragments to general page
Blunt-end cloning
cDNA cloning
Site-directed mutagenesis
Mutation genotyping (SNP analysis)

PCR products

All of the PCR products obtained using Advantage HD Polymerase Mix will possess blunt ends due to the 3' → 5' exonuclease activity of the enzyme. Consequently, the PCR products can be directly cloned into blunt-end vectors. If primers having sufficient vector sequence homology are used, then products can be cloned directly using In-Fusion technology.

Amplification with Advantage HD Polymerase using a single PCR cycling condition

Amplification with Advantage HD Polymerase using a single PCR cycling condition. 1–3 μl PCR products were run on a 1% agarose/EtBr gel. Lanes 2–7: Different sized fragments of the bovine pancreatic trypsin inhibitor (BPTI) gene were amplified from 100 ng of calf thymus genomic DNA. Lane 2: 500 bp. Lane 3: 900 bp. Lane 4: 1,500 bp. Lane 5: 2,000 bp. Lane 6: 2,500 bp. Lane 7: 3,500 bp. Lane 8: Amplification of 414 bp c-jun fragment from 100 ng human genomic DNA. Lane 9: Amplification of a ~3,000 bp lambda fragment from 1 ng lambda-gt10 lysate. Lane 1: Mixture (1:1) of lambda-Hind III and phi-X174-Hae III digests.

Outstanding fidelity of Advantage HD DNA Polymerase

Outstanding fidelity of Advantage HD DNA Polymerase. The error rate (mutation frequency) for Advantage HD Polymerase is lower than that of a number of commonly used DNA polymerases. Eight arbitrarily selected GC-rich regions from Thermus thermophilus HB8 genomic DNA were amplified with Advantage HD Polymerase and other polymerases. PCR products (approximately 500 bp representing each region) were cloned into suitable plasmids. Multiple clones were selected and subjected to sequence analysis to determine the error rate percentage (e.g., 1 error/100,000 bp = 0.001%).