Source: Purified from an E.coli strain carrying Taq-DNA-Polymerase overproducing plasmid. The original enzyme has been isolated from Thermus aquaticus. HotStart Taq-DNA-Polymerase is a modified form which is inactive at ambient temperature, having no polymerase activity. It is activated by a 15 minutes incubation at 95 - 97°C. This prevents extension of non-specifically annealed primers and of primer-dimers formed at low temperatures during PCR setup.
Associated activities: HotStart Taq-DNA-Polymerase is a highly processive 5’ --> 3’-DNA-polymerase with 5’ --> 3’-exonuclease activity. 3’ --> 5’-exonuclease activity lacks completely. Additionally, the enzyme adds nucleotides (mostly adenosines) to the 3´-ends of the DNA, so that TA-cloning is possible without further modifications.
Application & Quality control:
Primer extension reaction: the enzyme is free of nicking and primer extension activities as well as of exonucleases and unspecific endonucleases. SDS/PAGE: 95 kD-band, purity >98%. Activity and stability tested via PCR. The error rate per nucleotide per cycle is ~8,3x10 -5; the accuracy ~1,2x10 4. Estimated half life at 95ºC: 90 min.
Amount: 500 Units, Concentration: 5µl/U, Volume: 100 µl
incl. buffer set