TAQ DNA POLYM. 5 UNITS/UL (20X250 U)

General description

Taq DNA Polymerase is a highly processive 5′→3′ DNA polymerase that lacks 3′→5′ exonuclease activity. It is a single polypeptide chain with a molecular weight of approximately 95 kDa.
Taq DNA Polymerase was originally isolated from the thermophilic eubacterium Thermus aquaticus BM, a strain lacking Taq I restriction endonuclease. The enzyme was cloned in E.coli.

Application

Taq DNA Polymerase can be used in simple, routine PCR. Roche Applied Science Taq DNA polymerase is held to rigorous purity and quality standards. This preparation of recombinant Taq DNA Polymerase can be applied for:

• PCR
• RT-PCR
• qPCR
• Other primer-extension reactions, such as sequencing and labeling

Features and Benefits

Reliable reproducible results:
High lot-to-lot consistency.
No need to test each lot:
Taq DNA Polymerase is rigorously tested.
Prevent PCR carryover:
dUTP incorporation combination with Uracil-DNA Glycosylase prevents PCR cross-contamination.

Packaging

1 kit containing 2 components

Quality

Routine assays have medium size amplicons and 50% GC content. Taq DNA Polymerase has no proofreading or hot start features. It is optimally active at +75°C and pH 9.
Lack of restriction endonuclease:
The enzyme originally isolated from T. aquaticus BM lacks Taq I restriction endonuclease activity.Each lot is PCR tested using λDNA. Each lot is also tested for the absence of exo- and endonucleases, and nicking activities according to the current Quality Control procedures.

Unit Definition

One unit Taq DNA Polymerase is defined as the amount of enzyme that incorporates 10 nmol of total deoxyribonucleosidetriphosphates into acid precipitable DNA within 60 min at +65 °C under the assay conditions given above.

Unit Assay: Incubation buffer:
67 mM Tris/HCl; pH 8.3/25 °C, 5 mM MgCl₂, 10 mM Mercaptoethanol, 0.2% Polydocanol, 0.2 mg/ml Gelatine, 0.2 mM each dATP, dGTP, dTTP and 0.1 mM dCTP.

Incubation procedure:
M13mp9ss, M13 primer (17mer) and 1 μCi (α^-32P) dCTP are incubated with suitable dilutions of Taq DNA Polymerase in 50 μl incubation buffer at +65 °C for 60 minutes. The amount of incorporated dNTPs is determined by trichloroacetic acid precipitation.

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.