TnATaq DNA polymerase

TnATaq  DNA  polymerase is a thermostable enzyme isolated from E. coli which encodes Taq DNA polymerase gene. This enzyme contains 5’-3’ poly merase and 5’-3’ exonuclease activity.

50mM Tris-HCl pH7.9, 50mM KCl, 0.1mM EDTA,

1mM DTT, 0.5mM PMSF , 50% glycerol.

Containing 15mM MgCl₂

One unit is defined as the amount of enzyme that will incorporate 10nmole of dNTP into acid-insoluble material in 30 minutes at 74ºC. The reaction conditions are: 50mM Tris-HCl pH8.8, 50mM NaCl, 5mM MgCl₂, 200uM each of dATP, dCTP, dGTP, dTTP, 10 mg activated calf thymus DNA and 0.1mg/ml BSA in a final volume of 50 μl.

50% glycerol (v/v), 20 mM Tris-HCl pH 8.7 at -20ºC, 100 mM KCl, 0.1 mM EDTA.

E coli clone

The enzyme is free of nicking and priming activities, exonucleases and non-specific endonucleases. SDS/PAGE – 95 kD band. Activity and stability tested via thermo-cycling. The error rate per nucleotide per cycle is ~ 2.5 x 10-5; the accuracy is ~ 4 x 10-4. Estimated half life at 95ºC is 0.5 hours.

• Amplification of DNA fragments up to 5 kb
• Labelling of PCR products with modified nucleotides (biotin-dUTP, fluorescein-dUTP)
• Cycle sequencing
• Generation of PCR product for TA cloning

IMPORTANT: Annealing temperature should be 2-6ºC lower than the primer melting temperature.

Shipping and temporary storage at -20 and for up to 1 month at room temperature has no detrimental effects on the quality of TnATaq DNA polymerase.