- Производитель:
- Roche
Enzyme Commission (EC) Number: | 2.7.7.7 ( BRENDA | IUBMB ) |
Кат. номер |
4340850001 |
4340868001 |
Pwo SuperYield DNA Polymerase exhibits increased thermal stability with a half life of greater than 2 hours at +100 °C compared to Taq DNA Polymerase with a half life of less than 5 min at this temperature.
Pwo SuperYield DNA Polymerase, originally isolated from Pyrococcus woesei, is a processive 5 3 DNA polymerase with 3 5 exonuclease proofreading activity; 5 3 exonuclease activity is not detectable.
The enzyme accepts modified nucleotides for efficient labeling of nucleic acids by PCR.
PCR products are blunt-ended directly useable for blunt-end ligation.
Using the magnesium-containing reaction buffer supplied, the final MgCl2 concentration is 1.5mM.
Specificity
Star activity: In cases where star activity is observed and/or the activity of the enzyme in the PCR mix is low, first purify the amplification product prior to the restriction enzyme digest using High Pure PCR Product Purification Kit.
- Pwo Super Yield DNA Polymerase combines the recombinant enzyme Pwo DNA Polymerase with a newly optimized buffer system. Pwo SuperYield DNA Polymerase is used for the amplification of DNA with the intent to sequence the amplification product or to clone the product (e.g., for the expression of the gene product). The high fidelity of this enzyme makes it particularly suitable for: High fidelity PCR
- Site-directed mutagenesis
- Cloning
- Gene expression
- Study of allelic polymorphism in individual RNA transcripts
- Characterization of rare mutations in tissue
- Characterization of the allelic stage of single cells or single DNA molecules
Features and Benefits
Pwo SuperYield DNA Polymerase yields more high fidelity PCR product. The optimized buffer delivers superior results. Amplify fragments up to 3kb. A GC-RICH Solution is included for difficult templates.
- Higher yield and 18-fold higher fidelity
- High performance with difficult templates.
- Reduce working steps in cloning.
Packaging
1 kit containing 3 components
Quality
Each lot is assayed using activated DNA. PCR testing used λ and human genomic DNA.
Unit Definition
One unit Pwo SuperYield DNA Polymerase is defined as the amount of enzyme that catalyzes the incorporation of 10 nmol total deoxynucleoside triphosphates into acid precipitable DNA within 30 minutes at +70 °C.
Unit Assay: Incubation buffer for assay on activated DNA
20 mM Tris-HCl, pH 8.8 (20 °C), 50 mM KCl, 2.5 mM MgCl2, 10 mM 2-mercaptoethanol, 0.2 mM of each dATP, dCTP, dGTP, dTTP.
Incubation procedure
12.5 mg activated calf thymus DNA and 0.1 mCi [-32P]dCTP are incubated with 0.01 to 0.1 U Pwo SuperYield DNA Polymerase in 50 µl incubation buffer with a paraffin-oil overlay at +70 °C for 30 minutes. The amount of incorporated dNTPs is determined by trichloroacetic acid precipitation followed by scintillation counting.
Volume Activity: 5 U/µl
Pwo DNA Polymerase accepts modified nucleotides like digoxigenin-dUTP, biotin-dUTP, or fluorescein-dUTP. Thus, it can add these nucleotides to DNA during PCR. These nonradioactively labeled products can be used as a hybridization probe in many applications.
For life science research only. Not for use in diagnostic procedures.
usage | sufficient for ≤200 reactions (04340850001), sufficient for ≤40 reactions (04340868001) |
Quality Level | 100, |
feature | Difficult Templates/Specialty Enzymes PCR, dNTPs included: no, hotstart: no |
packaging | pkg of 100 U (04340868001), pkg of 500 U (04340850001 [2 x 250 U]) |
Торговая марка | Roche |
parameter | 72 °C optimum reaction temp. |
application(s) | PCR: suitable |
shipped in | dry ice |
storage temp. | −20°C |
RIDADR | NONH for all modes of transport |
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