PrimeSTAR HS DNA Polymerase
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 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.
The fidelity of PrimeSTAR HS DNA Polymerase was calculated by direct sequencing analysis to provide a more realistic estimate of the rate of incorporated errors compared with other commonly used indirect phenotypic analyses such as lacI. Calculation methods such as lacI, which are often used by other enzyme suppliers, are prone to error due to inherent lethal and silent mutations.
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.
For the most challenging PCR applications, enhanced versions of PrimeSTAR HS are available with the ability to amplify GC-rich sequences easily, even higher fidelity, and for long-range PCR:
- PrimeSTAR HS with GC buffer provides reliable, high-fidelity amplification for the most difficult GC-rich content (75% or greater).
- PrimeSTAR Max products provide the highest fidelity and fastest extension time of any Takara Bio product.
- PrimeSTAR GXL products provide high fidelity plus the ability to amplify long and GC- or AT-rich targets.
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
More Information
Applications
- High-fidelity PCR
- Amplification of cDNA libraries
- cDNA cloning for protein expression
- Site-directed mutagenesis and mutant genotyping (e.g., SNP analysis)
Note regarding fidelity comparison
We use sequencing analysis to determine incorporation error rate because most researchers who require high-fidelity amplification perform the following: PCR, cloning, and sequencing. Determination of error rate by direct sequence analysis, therefore, corresponds closely to the method that most researchers would use in their own applications. Please note that values obtained using different fidelity measurement methods (e.g., direct sequencing, Kunkel method, Cline method, lacI method) do not yield directly comparable data.
Purity
Neither nicking, endonuclease, nor exonuclease activity are detected following incubation of 0.6 µg of supercoiled pBR322 DNA or lambda DNA with 10 units of enzyme for 1 hour at 74°C.
Storage
Shipped and stored at –20°C. Avoid repeated freeze-thaw and vigorous stirring. Once thawed, dispense into PCR tubes and store at –20°C.
Performance
Enzymatic performance was confirmed by single-fragment PCR using both lambda DNA (amplified fragments: 8, 10, 12, 15 kb) and human genomic DNA (amplified fragments: 0.5, 1, 2, 4, 6, 8 kb) as templates.
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.
Bulk, custom, and OEM information
If you are interested in bulk purchasing, custom packaging, custom formulations (including glycerol-free and high concentration), or partnership opportunities, please contact Corporate Development at bd_oem@takarabio.com to discuss your needs or visit our OEM page to submit an inquiry.
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
See what our customers are saying about PrimeSTAR HS DNA Polymerase!
"The activity of primeSTAR HS DNA polymerase is very good. Thank you for your sample! We will use your enzyme and recommend to my colleagues."
—Shigui Zhu, PROTEIN ONE
"This polymerase solved the problems I was having both with PCR and sub-cloning with the products."
—Ethan Duane, UNIV OF NEW HAMPSHIRE
Takara Bio USA, Inc.
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