Description
General description
Contents
Taq DNA Polymerase, 5 U/μl
PCR Buffer, 10x concentrated, with MgCl₂
PCR Nucleotide Mix
For maximum convenience, select the ready-to-use 2x concentrated PCR Master.
Each dNTPack contains 10 mM additive-free sodium salt nucleotides as a ready-to-use mix.
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 preparation obtained from E.coli is free of nonspecific engo- or exonucleases according to the current quality control procedures.
Application
Achieve the most consistent results in simple, routine PCR by using Roche Applied Science Taq DNA Polymerase, which is held to our rigorous purity and quality standards. The Taq DNA Polymerase, dNTPack, combines our standard preparation (5 U/μl) of recombinant Taq DNA Polymerase with our convenient, ready-to-use PCR Nucleotide Mix. In all other respects, this preparation is identical to our standard preparation and can be used for:
PCR
RT-PCR
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 3 components
Preparation Note
Containers which are opened must be carefully resealed and
kept upright to prevent leakage
Analysis Note
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.
Other Notes
For life science research only. Not for use in diagnostic procedures.
One unit Taq DNA Polymerase is defined as the amount of enzyme that incorporates 10 nmol of total deoxyribonucleosidetriphosphates into acid precipitable DNA within 30 minutes at +75 °C under the assay conditions stated under unit assay.
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
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.
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.
