PrimeSTAR HS DNA Polymerase

Brand	Takara Bio

PrimeSTAR HS DNA Polymerase is a novel high-fidelity DNA polymerase that allows high-efficiency amplification of large DNA products (up to 8.5 kb for human genomic DNA; up to 22 kb for lambda DNA). Its excellent performance is achieved by superior proofreading ability due to a robust 3′ → 5′ exonuclease activity. The antibody-mediated hot-start (HS) feature provides improved specificity, thus reducing background and proving useful during highly demanding cloning and sequencing applications. PrimeSTAR HS is available in flexible and convenient formats:

Individual Components—PrimeSTAR HS DNA polymerase is supplied with a tube of 5X PrimeSTAR Buffer (with Mg2+) and a tube of dNTPs.
2X PCR master mix—a premix that includes PrimeSTAR HS DNA Polymerase, reaction buffer, and dNTPs for simple and convenient PCR setup and minimal pipetting steps.

PrimeSTAR HS DNA Polymerase also offers excellent amplification efficiency and a fast reaction time due to its extremely high priming efficiency. Highly specific amplifications can be achieved using short annealing times of just 5–15 seconds.

Overview

High accuracy and strong exonuclease activity, resulting in an extremely low error rate of only 12 errors per 250 kb
Higher amplification efficiency compared with standard Taq polymerase
Excellent performance even with GC-rich templates
High tolerance to varying reaction conditions (a single PCR cycling protocol can be used to amplify products of varying sizes)
Amplification of targets up to 8.5 kb with human genomic DNA, 10 kb with E. coli genomic DNA, and 22 kb with lambda DNA
Fast reaction time due to increased priming efficiency
Hot-start PCR enzyme prevents false initiation events during reaction assembly due to mispriming or primer digestion

Applications

High-fidelity PCR
Amplification of cDNA libraries
cDNA cloning for protein expression
Site-directed mutagenesis and mutant genotyping (e.g., SNP analysis)

PCR products

A significant percentage of PCR products obtained using PrimeSTAR HS DNA polymerase will have blunt ends. As a result, PCR products can be directly cloned into blunt-ended vectors. If necessary, phosphorylate PCR products before cloning

PrimeSTAR High Amplification Efficiency

PrimeSTAR High Amplification Efficiency. Amplification efficiency was compared using high fidelity enzymes from Company A and Company B. (Target: Human DCLRE1A gene [2kb]) Reaction mixtures were prepared and PCR cycling conditions were performed according to each company’s protocol (50 µL PCR reaction). These results demonstrate that PrimeSTAR provides excellent amplification efficiency with higher specificity than other suppliers’ high fidelity enzymes. In addition, the detection sensitivity was higher by one order of magnitude.

Sequence analysis of amplified DNA fragments

Sequence analysis of amplified DNA fragments. Sequence analysis is the most accurate method to investigate mutation frequency. This analysis revealed that PrimeSTAR HS has higher fidelity than an alternative high fidelity enzyme from Company A, with only 12 mismatched bases per 249,941 total bases, and 10X higher fidelity than Taq DNA polymerase. These results demonstrate PrimeSTAR HS DNA Polymerase’s suitability for PCR amplifications that require extreme accuracy.

PrimeSTAR HS DNA Polymerase

Additional information