04743750001

Pwo SuperYield DNA Polymerase, dNTPack

sufficient for ≤40 reactions, pkg of 100 U, optimum reaction temp. 72 °C

• Synonym(s): PCR | Polymerase
• Enzyme Commission number: 2.7.7.7 (BRENDA, IUBMB)
• UNSPSC Code: 41106300

Properties

• usage: sufficient for ≤40 reactions
• packaging: pkg of 100 U
• manufacturer/tradename: Roche
• parameter: 72 °C optimum reaction temp.
• storage temp.: −20°C (−15°C to −25°C)

Description

General description

Pwo SuperYield DNA Polymerase combines the recombinant enzyme Pwo DNA Polymerase with an optimized buffer system. This buffer system enhances the enzymatic properties of the polymerase, resulting in higher yields of the amplification reaction without changing the fidelity of DNA synthesis. This enzyme delivers excellent results due to its enzyme design and optimized buffer system. It amplifies fragments up to 3 kb - even longer amplicons are possible from simple templates.
Pwo 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 1.5 mM.

Specificity

Star activity: In cases where star activity is observed and/or the activity of the enzyme in the PCR mix is low, purify the amplification product prior to the restriction enzyme digest using High Pure PCR Product Purification Kit.

Application

Pwo SuperYield 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 3 kb. A GC-RICH Solution is included for difficult templates. Each dNTPack contains the 10 mM additive-free sodium salt nucleotides in a ready-to-use mix.

• Higher yield and 18-fold higher fidelity

compared to Taq DNA Polymerase.

• High performance with difficult templates.

The GC-RICH Solution is for GC-rich PCR.

• Reduce working steps in cloning.

Perform digests directly in Pwo SuperYield PCR mix.

• Cost-effective.

Use the convenient premixed solution of PCR grade dNTPs.

Packaging

1 kit containing 4 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 [α-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

Other Notes

For life science research only. Not for use in diagnostic procedures.

Legal Information

Use of this product is covered by US patent claims and corresponding patent claims outside the US. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patent claims require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Kit Components Only

• Pwo SuperYield DNA Polymerase 5 U/μl
• PCR Buffer, containing 15 mM MgSO₄ 10x concentrated
• GC-RICH Solution 5x concentrated
• PCR Nucleotide Mix

Safety Information

• Hazard Statements: H412
• Precautionary Statements: P273 - P501
• Hazard Classifications: Aquatic Chronic 3
• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 2
• Flash Point(F): does not flash
• Flash Point(C): does not flash