Taq DNA polymerase was isolated and purified from E.coli with a molecular weight of 94 KD. It has 5 \'- 3\' DNA polymerase activity and 5 \'- 3\' exonuclease activity, but no 3 \'- 5\' exonuclease activity. In the PCR reaction, Taq DNA polymerase was extended at a rate of 1-2 KB / min, and the product had a 3 \'end band, which could be directly used for t / a vector cloning. The enzyme is stable and specific without contamination of exogenous nuclease and bacterial genomic DNA.Activity definition 1 unit (U) Taq DNA polymerase activity was defined as the amount of enzyme required to add 10 nmol of deoxynucleotides into acid insoluble substances using the activated salmon sperm DNA as template at 74 ƒ for 30 minutes.
Quality Control
The purity of Taq DNA polymerase detected by SDS-PAGE was more than 99%, and no exogenous nuclease activity was detected by detection; there was no host residual DNA detected by PCR method; single copy gene in human genome could be amplified effectively; no obvious activity change was found after storage at room temperature for one week.
Enzyme storage buffer
20 mM Tris-HCl (pH 8.0);0.1 mMEDTA;1 mMDTT;100 mMKCl ; Stabilizers; 50% glycerol 10×Taq Buffer
200mm Tris HCl (pH 8.4); 200mm KCl; 15mm MgCl2; other ingredients.
Scope of application
It is generally used for PCR amplification of DNA fragments, DNA labeling, primer extension, sequencing, and addition of a at the flat end. The product can be directly used for t / a vector cloning.
matters needing attention
When taking Taq DNA ploymerase for PCR reaction, please use the sterilized tip. |
