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Taq DNA Polymerase (with dNTPs)
Background:
Thermus aquaticus DNA polymerase (Taq DNA polymerase) was expressed in E. coli in large quantities and highly purified. The enzyme has thermostable DNA polymerase activity and the MW is 94 kDa.. This enzyme is suitable for PCR reactions; capable of amplifying DNA with various primers.
Application:
1) High-throughput PCR
2) Colony PCR
3) Incorporation of dUTP, dITP, and fluorescence-labeled
nucleotides
4) Primer extension
5) Addition of a single nucleotide (adenosine) at the
3’-blunt ends
General composition of PCR reaction mixture (total 50µl)
| Taq DNA polymerase (5 units/μl) | *0.25 μl |
| 10 x Reaction Buffer (Taq) | 5 μl |
| 2.5 mM (each) dNTPs | 4 μl |
| Template | <500ng |
| Primer 1 | 0.2～1.0μM (final conc.) |
| Primer 2 | 0.2～1.0μM (final conc.) |
| Sterile distilled water | up to 50μl |
*Use of excess amount is not recommended
SDS-PAGE of
Taq DNA
polymerase
PCR condition
98°C 10sec
57°C 30sec 25cycles
72°C 8min
(2min in the case of 2kb DNA)
