Taq DNA Polymerase (recombinant), Low Concentration
Detailní popis
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 |
|
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 |
|
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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