PCR Master Mix (2X)

PCR Master Mix (2X)
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Highly thermostable DNA polymerase for routine PCR.

. Detailní informace

Cena s DPH 2 819,30 Kč
Cena bez DPH 2 330,00 Kč
 200 react
Dostupnost Skladem
Kód produktu K0171

Detailní popis PCR Master Mix (2X)

Thermo Scientific Taq DNA Polymerase is a highly thermostable DNA polymerase from the thermophilic bacterium Thermus aquaticus. The enzyme catalyzes 5’→3’ synthesis of DNA, has no detectable 3’→5’ exonuclease (proofreading) activity and possesses low 5’→3’ exonuclease activity. In addition, Taq DNA Polymerase exhibits deoxynucleotidyl transferase activity, which frequently results in the addition of extra adenines at the 3’-end of PCR products. Recombinant Taq DNA Polymerase is the ideal tool for standard PCR of templates 5 kb or shorter.

PCR Master Mix is a 2X concentrated solution of Taq DNA Polymerase, dNTPs, and all of the components required for PCR, except DNA template and primers. This pre-mixed formulation saves time and reduces contamination due to a reduced number of pipetting steps required for PCR set up. The mix is optimized for efficient and reproducible PCR.

Highlights

  • Thermostable – half life is more than 40 min at 95°C
  • Generates PCR products with 3’-dA overhangs
  • Supplied with two buffers – 10X Taq Buffer with KCl and 10X Taq Buffer with (NH4)2SO4. The latter allows for PCR at wide range of magnesium concentrations and decreases unspecific priming
  • Incorporates modified nucleotides (e.g., biotin-, digoxigenin-, fluorescently-labeled nucleotides)

Applications

  • Routine PCR amplification of DNA fragments up to 5 kb
  • High throughput PCR
  • DNA labeling (see Reference 1,2,3)

Includes

Taq DNA Polymerase (recombinant):

  • Taq DNA Polymerase 5 U/µL
  • 10X Taq Buffer with KCl
  • 10X Taq Buffer with (NH4)2SO4
  • 25 mM MgCl2

Taq DNA Polymerase (recombinant), LC:

  • Taq DNA Polymerase 1 U/µL
  • 10X Taq Buffer with KCl
  • 10X Taq Buffer with (NH4)2SO4
  • 25 mM MgCl2

PCR Master Mix (2X):

  • Taq DNA polymerase (0.05 U/µL), reaction buffer, 4 mM MgCl2, and 0.4 mM of each dNTP
  • Nuclease-free water

Note

  • The error rate of Taq DNA Polymerase in PCR is 2.2 x 10-5 errors per nt per cycle, as determined by a modified method described in (see Reference 4). Accordingly, the accuracy of PCR is 4.5 x 104. Accuracy is an inverse of the error rate and shows an average number of correct nucleotides incorporated before an error occurs.
  • The 10X Taq Buffer without Detergent is recommended for microarray experiments.
10X Taq Buffer with (NH4)2SO4 750 mM Tris-HCl (pH 8.8 at 25°C), 200 mM (NH4)2SO4, 0.1% (v/v) Tween 20.
10X Taq Buffer with KCl 100 mM Tris-HCl (pH 8.8 at 25°C), 500 mM KCl, 0.8% (v/v) Nonidet P40.
Definition of Activity Unit
  • One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30 min at 70°C.
  • Enzyme activity is assayed in the following mixture: 67 mM Tris-HCl(pH 8.8 at 25°C), 6.7 mM MgCl2, 1 mM 2-mercaptoethanol, 50 mM NaCl, 0.1 mg/mL BSA, 0.75 mM activated calf thymus DNA, 0.2 mM of each dNTP, 0.4 MBq/mL [3H]-dTTP.
Hazardous No
Inactivation Inactivated by phenol/chloroform extraction.
Inhibition Inhibitors: ionic detergents (deoxycholate, sarkosyl and SDS) at concentrations higher than 0.06, 0.02 and 0.01%, respectively (5).
Molecular Weight 94 kDa monomer.
Quality Control
  • The absence of endo-, exodeoxyribonucleases, and ribonucleases confirmed by appropriate quality tests.
  • Functionally tested in PCR.
Source E. coli cells with a cloned pol gene from Thermus aquaticus YT1.
Storage Buffer The enzyme is supplied in 20 mM Tris-HCl (pH 8.0), 1 mM DTT, 0.1 mM EDTA, 100 mM KCl, 0.5% (v/v) Nonidet P40, 0.5% (v/v) Tween 20, and 50% (v/v) glycerol.
Storage Condition -20 C

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