Hot Start Taq DNA Polymerase is a hot-start Taq DNA Polymerase modified by anti-TAQ monoclonal antibody. The anti-TAQ monoclonal antibody binds to Taq enzyme before high temperature heating, inhibiting the activity of the polymerase. The nonspecific amplification caused by nonspecific annealing of primer or primer dimer at low temperature is inhibited. Anti-taq monoclonal antibodies are denatured in the initial DNA denaturing step of the PCR reaction, so they can be used under conventional PCR reaction conditions without special inactivation treatment. In qPCR reaction, it can significantly improve the amplification efficiency of fluorescent PCR (especially for low copy number templates), improve the perfection of its amplification curve, and it has good stability, high repeatability and strong specificity. When the enzyme was left at room temperature for a week, the enzyme activity remained above 95%.
Activity definition: 1 unit (U) Taq DNA Polymerase activity is defined as the amount of enzyme required to incorporate 10 nmol of deoxynucleotides into an acid-insoluble substance using activated salmon sperm DNA as a template at 74℃for 30 minutes.
Quality control: The purity of Taq DNA Polymerase detected by SDS-PAGE was more than 99%. No exogenous nuclease activity was detected. No host residual DNA was detected by PCR. Can effectively amplify single copy genes in the human genome; Stored at room temperature for one week, no significant change in activity.
Enzyme storage buffer: 20mM Tris-HCl (pH 8.0); 0.1 mM EDTA; 1 mM DTT; 100 mM KCl ; 50% glycerol; stabilizer
Scope of application: It can be used for PCR amplification, DNA labeling, primer extension, sequence determination, flat-end addition of A, etc. of DNA fragments less than 6kb. The product can be directly used for T/A vector cloning.
Recommended PCR conditions: the pre-denaturation temperature is 95 degrees, the time is 5-10 minutes, and other reaction conditions are the same as that of ordinary Taq enzymes.
PCR system (50μl reaction system as an example)
Template < 0.5 mu g
Upstream primer (10μM) 1μl
Downstream primer (10μM) 1μl
10×Buffer (including Mg2+) 5μl
dNTP (2.5mM each) 4μl
Heat activated Taq enzyme 0.5-1μl
ddH2O up to 50μl
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G8142 GoldView Type II Nucleic Acid Stain (5000×)
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