Taq DNA Polymerase (a 94 KDa thermostable DNA polymerase) can amplify target DNA up to 5 kb. It is more suitable to amplify the target template between 200 bp to 3000 bp, at elongation velocity of 0.9 to 1.2 kb per min. It has 5' to 3' polymerase activity with a little 3' to 5' exonuclease activity. Taq DNA Polymerase has deoxynucleotidyl transferase activity, which frequently results in the addition of extra adenine at the 3' end of PCR products.
One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nM of dNTPs into an acid-insoluble form in 30 minutes at 70℃ using sperm DNA as substrate.
10×Taq PCR Buffer (Mg2+ Plus) 5.0 μl 1×
2.5 mM dNTP mixture 4.0 μl 0.2 mM each
Primer 1 (10 pmol/μl) 2.0 μl 0.4 pmol/μl
Primer 2 (10 pmol/μl) 2.0 μl 0.4 pmol/μl
Template DNA* 2.0 μl 10 ng to 1.0 μg
Taq DNA Polymerase (5 U/μl) 0.5 μl 0.1 U/μl
Autoclaved distilled water Up to 50.0 μl
*DNA template: mammalian genomic DNA 0.1 to1 μg; E.coli genomic DNA 10 to100 ng; plasmid DNA 0.1 to 10 ng etc.
Incubate tube in a thermal cycler for 3 minutes at 95°C (complete denaturation of the template). Set the thermal cycler; Denature 95°C for 45 seconds; Anneal 55°C for 30 seconds; Extend 72°C for 1 minute, 30 seconds; Final elongation for an additional 10 minutes at 72°C. Perform 25-35 cycles of PCR amplification.
USE- Cat. no. 4200MX dNTP