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 into E. coli.

Request

Taq DNA polymerase can be used in simple and routine PCR. Roche Applied Science Taq DNA Polymerase is subject to rigorous standards for purity and quality. This recombinant Taq DNA polymerase preparation can be ordered for:
• PCR
• RT-PCR
• qPCR
• Other primer extension reactions, such as sequencing and labeling

Features and Benefits

• Reliable and reproducible results:
• High consistency from batch to batch.
• It is not necessary to test every batch:
• Taq DNA Polymerase is rigorously tested.
• Avoid PCR carryover:
• The combination of dUTP incorporation with uracil-DNA glycosylase prevents cross-contamination by PCR.

Packaging

1 kit containing 2 components

Quality

Routine assays have medium-size amplicons and a GC content of 50%. Taq DNA Polymerase does not have hot start or review functions. 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 tested by PCR using λDNA. Each lot is also tested for the absence of Exo and endonucleases, and cutting activities according to current quality control procedures.

Definition of unit

One unit of Taq DNA polymerase is defined as the amount of enzyme that incorporates 10 nmol of total deoxyribonucleoside triphosphates into acid precipitable DNA in 60 minutes 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% polidocanol, 0.2 mg / ml gelatin, each 0.2 mM dATP, dGTP, dTTP and 0.1 mM dCTP.

Incubation procedure:

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

Volume activity:

5 U / μl

Other notes

For life science research only. It should not be used in diagnostic procedures.

Legal information

The use of this product is covered by US patent applications and corresponding non-US patent applications. The purchase of this product includes limited and non-transferable immunity from a lawsuit under the claims of Previous patents for using only this amount of product for the buyer’s own internal research.

No right is expressly conveyed under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including but not limited to reporting the results of the buyer’s activities in exchange for a fee or other commercial consideration, by implication or by a legal impediment. This product is for research only. Diagnostic uses under Roche patent claims require a separate license from Roche. More information on purchasing licenses can be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Center Drive, Foster City, California 94404, USA.