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DESCRIPTION

Taq DNA polymerase is the most widely used thermostable DNA polymerase derived from the thermophilic bacteria Thermus aquaticus (Taq) YT-1. The enzyme possesses a 5’→3’ polymerase activity and a double-strand specific 5’→ 3’ exonuclease activity.

FEATURES

  • Tolerates various kinds of PCR protocols.
  • Applicable for hot start technology by adding anti-Taq antibody (Code No. TCP-101).
  • PCR products can be cloned by using a TA cloning method.
  • Incorporates dUTP, dITP, and fluorescently-labeled nucleotides.

APPLICATIONS

  • PCR
  • Primer extension

SOURCE

E. coli strain carrying the cloned Taq DNA polymerase gene from Thermus aquaticus (Taq) YT-1.

UNIT DEFINITION

One unit is defined as the amount of enzyme that will incorporate 10 nmoles of dNTP into an acid insoluble material in 30 min at 75ºC.

STORAGE CONDITION

20 mM Tris-HCl (pH 8.0), 100 mM KCl, 0.1 mM EDTA, 0.5% Nonidet® P-40, 0.5% Tween® 20, 50% Glycerol.
Store at -20ºC

COMPONENTS

This reagent includes the following components for 100-200 reactions;

rTaq DNA Polymerase (2.5U/µl)
10×Buffer
25 mM MgCl2
2mM dNTPs

TYPICAL PCR REACTION SETUP

Normal PCR

Component Volume Final Concentration
10x Buffer 5 µl
2mM dNTPs 5 µl 0.2 mM each
25mM MgCl2 3 µl 1.5 mM
10pmol/ul Primer #1 1.0 µl 0.2 µM
10pmol/ul Primer #2 1.0 µl 0.2 µM
Template DNA X µl Genomic DNA 10~1000 ng/50 µl
Plasmid DNA 1~50 ng/50 µl
cDNA ~200 ng (RNA equiv.)/50 µl
PCR grade water Y µl
Diluted rTth DNA polymerase (1.0U/µl) 1.25-2.5 µl 1.25-2.5 U / 50 µl
Total reaction volume 50 µl

PCR CYCLE CONDITIONS

*Extension time should be set at 1 min per 1 kb of target length.

APPLICATION DATA

Example 1.Amplification of 180 bp–1.3 kb genes from human genomic DNA

Distinct and specific amplified bands from 180 bp to 1.3 kb were observed with rTaq DNA polymerase by 1% agarose gel electrophoresis.

REFERENCES

  • F.C. Lawyer, S. Stoffel, R.K. Saiki, K. Myambo, R. Drummond, D.H. Gelfand., J. Biol. Chem., 264: 6427-6437 (1989)
  • T. Nagahama, K. Sugiura, S. Lee, H. Morita, Y. Adachi, A.H. Kwon, Y. Kamiyama, S. Ikehara, Stem cells, 19: 425-435 (2001)