Sigma-Aldrich Taq DNA Polymerase from Thermus aquaticus

General description

Taq polymerase is a thermostable DNA polymerase named after the thermophilic bacterium Thermus aquaticus. A stable deoxyribonucleic acid (DNA) polymerase (with a temperature optimum of 80°C) has been purified from the extreme thermophile Thermus aquaticus. The enzyme is free from phosphomonoesterase, phosphodiesterase and single-stranded exonuclease activities. Maximal activity of the enzyme needs all four deoxyribonucleotides and activated calf thymus DNA.[1]

Application

Taq DNA Polymerase from Thermus aquaticus has been used:

• in the process of DNA extraction (during gene amplification and sequencing)
• in genotyping[2]
• in polymerase chain reaction (PCR) to study the constitutive production of epithelial neutrophil activating peptide 78 (ENA-78) and interleukin-8 (IL-8)[3]
• for amplification of RNA from primary endothelial cells by conventional PCR[4]

Biochem/physiol Actions

Taq polymerase catalyzes oligonucleotide primer-driven, DNA template dependent incorporation of dNTPs into complimentary DNA strands. It displays both 5′ to 3′ polymerase and exonuclease activities.

Features and Benefits

• MgCl2 provided in a separate tube to allow MgCl2 optimization
• Can withstand repeated heating to 95 °C without significant loss of activity

Packaging

Taq DNA Polymerase with 10× reaction buffer without MgCl2. Includes a separate tube of 25 mM MgCl2Taq DNA polymerase comes with the choice of an optimized 10× reaction buffer including MgCl2 (D1806) or a 10× reaction buffer without MgCl2 plus a separate tube of MgCl2 for titration (D4545). The latter option may be necessary to determine optimal conditions for amplification.

Other Notes

Taq DNA Polymerase is a specialized thermostable enzyme isolated from the thermophilic bacterium Thermus aquaticus. The recombinant form of this enzyme is expressed in E. coli. This 94 kDa protein shows no detectable levels of contaminating endonucleases or exonucleases by SDS-PAGE. It has both 5′→3′ polymerase and exonuclease activity.

Unit Definition

One unit will incorporate 10 nmol of total dNTPs into acid-precipitable DNA in 30 min at 74 °C.