Taq DNA polymerase (Standard buffer)
Thermus aquaticus DNA polymerase (Taq DNA polymerase) gene was expressed in E.coli in large quantities and highly purified. The enzyme has thermostable DNA polymerase activity and the MW is 94 kDa, same as that of the natural enzyme.
This enzyme is suitable for PCR reactions; capable of amplifying DNA with various primers.
- High-throughput PCR
- Colony PCR
- Incorporation of dUTP, dITP, and fluorescence-labeled nucleotides
- Primer extension
- Addition of a single nucleotide (adenosine) at the 3’-blunt ends
General composition of PCR reaction mixture (total 50ul)
|Taq DNA polymerase (5 units/ul)||*0.25 ul|
|10 x Standard Buffer (Taq)||5 ul|
|2.5mM (each) dNTPs||4ul|
|Primer 1||0.2～1.0uM (final conc.)|
|Primer 2||0.2～1.0uM (final conc.)|
|Sterile distilled water||up to 50ul|
*Use of excess amount of enzyme is not recommended.
20mM Tris-HCl (pH 8.0), 100mM KCl, 0.1mM EDTA, 1mM DTT, 50% glycerol, 0.5% Tween20, 0.5% Igepal CA-630, Store at -20℃
5 units/ul, where one unit is defined as the amount of enzyme that can incorporate 10 nmols of total dNTPs into an acid-insoluble material in 30 minutes at 74℃ when activated salmon sperm DNA was used as template/primer.
Greater than 95% of protein determined by SDS-PAGE (CBB staining) (Fig.1) The absence of endonucleases and exonucleases was confirmed.
PCR Test: Good amplification result was obtained in PCR reaction using λDNA as a template (Fig.2).