PCR amplification Kit with HS-Taq (+MgCl2)

Features
The mix contains all reagents necessary for amplification of genomic DNA, cDNA and cloned DNA templates including hot-start Taq DNA polymerase and 5× PCR buffer with MgCl2
Application
  • Hot-start PCR
  • High-throughput PCR
  • Conventional PCR with high reproducibility
  • Generation of PCR products for TA cloning
  • RT-PCR (two-step method)

Basic description

BioMaster HS-Taq PCR Kit (+MgCl2) contains recombinant HS-Taq DNA polymerase and solutions of every component needed for performing conventional PCR under hot-start conditions, except for DNA template and primers. The mix includes: HS-Taq DNA polymerase solution (5 U/µl), 5× PCR buffer (+MgCl2), 50 mM MgCl2, 50× dNTP mix and 6× loading buffer. All reagents are of high quality and optimized for PCR.

HS-Taq DNA polymerase is a recombinant Taq DNA polymerase inactivated by specific monoclonal antibodies. HS-Taq DNA polymerase remains inactive at temperatures lower than 70 °С, which allows to avoid the formation of non-specific products and primer dimers at low temperatures during preliminary steps. The activation of HS-Taq DNA polymerase is achieved during the first cycle of PCR amplification after short incubation at 95 °С for 5 min. Recombinant Taq DNA polymerase has 5´-3´ DNA-dependent DNA polymerase activity and 5´-3´ exonuclease activity of native Taq DNA polymerase from Thermus aquaticus. The extension rate of Taq DNA polymerase depends on the complexity of DNA template and is approximately 1 kbp/min. Recombinant Taq DNA polymerase is ideal for conventional PCR of templates up to 5 kbp.

5× PCR buffer (+MgCl2) is optimized for performing efficient and reproducible PCR. The buffer contains MgCl2 (10 mM) and components increasing half-life and processivity of HS-Taq DNA polymerase by enhancing its stability during PCR. The buffer is chemically stable, inert and does not interfere with optimal annealing temperature or the parameters of template melting. Additional solutions of 50 mM MgCl2 and 50× dNTP mix included in the kit allow optimizing PCR conditions for individual primer pairs, while 6× loading buffer eases sample preparation for analysis and allows monitoring the progress of electrophoresis.

Areas of use

  • Classic PCR with "hot" start
  • Development of PCR products for TA cloning
  • RT-PCR one- and two-step

Benefits of using

  • Enzyme with hot-start capability increases specificity and sensitivity of PCR
  • HS-Taq DNA polymerase activation requires 5 min heating
  • Increased selectivity and reaction yield
  • PCR products can be further subjected to TA cloning due to the presence of deoxyadenosine overhangs in amplified DNA

Storage

1 year (under proper storage and transportation conditions).

Transportation

at -20 °С; not more than 50 thawing-freezing cycles.

Product passport

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