Taq DNA polymerase

概述:

Taq DNA polymerase is a highly thermostable DNA polymerase of Thermus aquaticus. The enzyme exhibits 5’→3’ polymerase activity and possesses 5’→3’ exonuclease activity but has no detectable 3’→5’ exonuclease activity. In addition, Taq DNA polymerase generates PCR products containing extra adenines at the 3’-ends. The resulting products can be used for TA cloning. Applications of this product include: routine PCR amplification of DNA fragments, generation of PCR products for TA cloning, DNA labelling and DNA sequencing.

参考文献:

1. Innis M A, et al: PCR protocols and applications: A laboratory manual, academic, New York, 1989.
2. Innis M A, et al: DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA, Proc. Natl. Acad. Sci. USA. 1988, 85:9436-9440.
3. Weyant R S, et al: Effect of ionic and nonionic detergents on the Taq polymerase. Biotechniques. 1990, 9:308-309.

存储稳定性:

Store product at –30oC to –15oC. For optimal performance aliquot product into smaller quantities after centrifugation and store at recommended temperature. Avoid repeated handling and multiple freeze/thaw cycles.

Product Description:

Recombinant Thermus aquaticus Taq DNA polymerase protein expressed in E. coli. cells.

Components:

Component Name
Taq DNA polymerase (5U/µl)
10X Taq buffer (Mg2+ plus)

Quality Control:

Exonuclease Activity: The product is tested in a reaction containing 10 U of Taq DNA Polymerase and 0.6 g of λ-Hind II DNA. After incubation at 37oC for 16 hours, there is no visually detectable change of pattern of DNA bands determined by agarose gel electrophoresis.

Endonuclease Activity: The product is tested in a reaction containing 10 U of Taq DNA Polymerase and 0.6 g of supercoiled pBR322. After incubation at 37oC for 4 hours, there is no visually detectable change of pattern of DNA bands determined by agarose gel electrophoresis.

Functional Assay: The human -1-antitrypsin gene is amplified for 30 cycles in a 50 l system using 1.25 U of Taq DNA Polymerase and 100 ng of human genomic DNA as template. A single DNA band of 360 bp is detected by 1% agarose gel electrophoresis.

Specific Activity:

The specific activity of Taq DNA polymerase was determined to be 5 Units/µl.
Unit Definition:
One unit (U) is defined as the amount of enzyme that incorporates 10 nmol of dNTPs into acid-insoluble products in 30 minutes at 74°C with activated salmon sperm DNA as template/primer.