Hzymes Taq DNA Polymerase is a thermophilic bacteria Thermus aquaticus-derived extremely thermostable DNA polymerase. The enzyme catalyzes 5”→3′ DNA synthesis, lacks 3”→5′ exonuclease activity, and has minimal 5”→3′ exonuclease activity. Furthermore, Taq DNA Polymerase is a deoxynucleotidyl transferase, which commonly results in the insertion of additional adenines at the 3′-end of PCR products. Recombinant Taq DNA Polymerase is an excellent instrument for standard PCR of templates of 5 kb or short.
Storage and transportation： Storage at -20℃ and transportation ≤ 0 ℃
Storage buffer: 10 mM Tris-HCl (pH 7.4 at 25 ℃), 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.5% Tween 20, 0.5% IGEPALCA-630 and 50% Glycerol.
Unit definition：One unit is defined as the amount of enzyme that will incorporate 15 nmol of dNTP into acid insoluble material in 30 minutes at 75°C.
Quality Control Assays
- Endonuclease Activity: Incubation of 20 U of enzyme with 4 μg pUC19 DNA for 4 hours at 37°C resulted in no detectable degradation of the DNA as determined by gel electrophoresis.
- 5 kb λDNA PCR: 25 Cycles of PCR amplification of 5 ng λDNA with 1.25 units of Taq DNA Polymerase in the presence of 200 µM dNTPs and 0.2 µM primers results in the expected 5 kb product.
- Exonuclease Activity: Incubation of a 50 µl reaction containing a minimum of 12.5 U of Taq DNA Polymerase with 10 nM 5´-FAM oligonucleotide for 30 minutes at 37℃ yields no detectable degradation.
- RNase Activity: Incubation of a 10 µl reaction containing 20 U of enzyme with 0.8 μg of MS2 RNA transcripts for 2 hours at 37°C resulted in no detectable degradation of the RNA as determined by gel electrophoresis.
Heat Inactivation: No
b ： The optimal concentration of Taq DNA Polymerase may range from 0.1 – 0.5 units/25 µL reaction in specialized applications.
Thermocycling Conditions for a Routine- PCR:
a: An initial denaturation of 1min at 95 °C is sufficient for most amplicons. For difficult templates, a longer denaturation of 2–3 min at 95°C is recommended. With colony PCR, an initial 5 min denaturation at 95°C is recommended.
b: The annealing step is typically 15-60 s. Annealing temperature is based on the Tm of the primer pair and is typically 45-68 ℃.