BioMaster Extended HS-Taq PCR kit contains recombinant Taq DNA polymerase, three reaction buffers and other components necessary for PCR, except for DNA template and primers. The kit includes recombinant HS-Taq DNA polymerase (5 U/µl); 10× PCR buffer; 5× Green buffer; 5× GC buffer; 50 mM MgCl2 (separate tube); 50× dNTP mix and 6× loading buffer.
HS-Taq DNA polymerase is a recombinant Taq DNA polymerase inactivated by specific monoclonal antibodies. HS-Taq DNA polymerase is inactive at temperatures up to 70 °С, which allows to avoid the formation of non-specific products and primer-dimers at low temperatures during preparation of PCR solution. The activation is achieved during 5-minute incubation at the first cycle of amplification at 95 °С. Recombinant Taq DNA polymerase has 5´-3´ DNA-dependent polymerase activity and 5´-3´ exonuclease activity of native Taq DNA polymerase form Thermus aquaticus. The rate of DNA synthesis by Taq DNA polymerase depends on the complexity of DNA template and is approximately 1 kbp/min. Recombinant form of the enzyme is ideal for conventional PCR of templates up to 5 kbp.
10× PCR buffer is optimized for most types of PCR including real-time PCR with intercalating dyes or fluorescent probes. The buffer is chemically stable, inert and does not affect the optimal temperature of primer annealing or characteristics of template melting.
5× Green buffer contains marker dyes (blue-green and yellow) and additives that increase solution density for easy gel loading. The buffer is developed for conventional PCR with further product yield assessment by gel electrophoresis. The buffer is chemically stable, inert and does not affect the optimal temperature of primer annealing or characteristics of template melting.
5× GC buffer is developed for amplification of GC-rich DNA regions and/or templates with complicated spatial structure. The buffer is chemically stable and contains substances that change primer annealing temperature and characteristics of template melting.
The buffers contain additives increasing half-life and processivity of HS-Taq DNA polymerase by enhancing its stability during PCR. Each buffer contains 1.5 mM MgCl2 (per 1× buffer solution). Additional tubes of 50 mM MgCl2 and 50× dNTP mix included in the kit allow easy optimization of PCR conditions for individual primer/template system.