Clontech’s Advantage HD Polymerase is a novel PCR enzyme that provides maximum fidelity as well as extended product length for High Definition PCR applications. The enzyme’s superior performance is due in part to the presence of a robust 3'–5' exonuclease activity as well as high priming efficiency. In addition, high specificity is achieved by inclusion of a hot start antibody that inhibits polymerase activity at ambient temperatures. When used with Clontech’s optimized reaction buffer, the Advantage HD Polymerase provides the high fidelity, sensitivity, and specificity required for critical applications such as gene cloning and amplification of cDNAs for library construction.
Superb Accuracy & Efficiency
The enzyme provides outstanding accuracy due to the presence of exonuclease activity that results in an extremely low error rate of just 12 errors per 250,000 bp. This degree of accuracy is superior to that of a number of commonly used DNA polymerases. In addition, amplification efficiency is greater than that of wild-type Taq Polymerase. The Advantage HD Polymerase also performs extremely well on GC-rich and other more complex templates.
Short Reaction Times
The high priming efficiency observed with Advantage HD translates into shorter annealing times and increased specificity. In addition, antibody-mediated hot start prevents false initiation events due to mispriming during reaction assembly.
The Advantage HD enzyme is active over a broad range of reaction conditions, and a single PCR cycling protocol can be used to efficiently amplify products of varying sizes. Consequently, Advantage HD Polymerase is an excellent choice for subcloning from cDNA libraries. The enzyme allows you to amplify targets of up to 8.5 kb from human genomic DNA, 10 kb from E. coli genomic DNA, and 22 kb from lambda DNA.
Advantage HD Polymerase is particularly useful for applications that require minimal errors, such as cDNA cloning, site-directed mutagenesis, and mutation genotyping (i.e., SNP analysis).
All of the PCR products obtained using Advantage HD Polymerase 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.
Each Advantage HD Polymerase kit provides enough enzyme and buffer for 200 25 µl reactions.