Hot Start Taq DNA Polymerase does not require special high temperature treatment. The pre-denaturation step in conventional PCR reaction conditions can start the activity of Taq enzyme. It is suitable for various Taq enzyme-based hot-start PCR and qPCR reactions. The PCR product has an A at the 3'end, which can be cloned directly with TA vector.
Unit definition At 74°C for 30 min, the amount of enzyme required to incorporate 10 nm dNTPs into the acid-insoluble precipitate is defined as 1 activity unit. Activity detection conditions: 50 mM Tris-Hcl (pH 9.0, 25°C), 50 mM NaCl, 5 mM MgCl2, 0.2 mM each dNTPs (including [3H]-Dttp), 200 μg/ml activated calf thymus DNA and 0.1 mg/ml BSA.
The purity detected by SDS-PAGE is greater than 99%. After detection of no exogenous
nuclease activity, PCR method detects no host DNA residues, which can effectively amplify
single-copy genes in the human genome.
PCR system components
- 1. Purity of template DNA: Many residual nucleic acid extraction reagents will affect the PCR
reaction, including protease, protein denaturant (such as SDS, guanidine salt), high
concentration salt (KAc, NaAc, sodium caprylate, etc.) and high concentration EDTA. The
amount of template with low purity (such as the template obtained by boiling method)
should not exceed 1/10 of the PCR reaction system (for example, the volume of template
added to the 50 μl reaction system should not exceed 5 μl). If the purity of the template DNA
is too poor, you can use our PCR product recovery kit (Cat. No. EP005) to purify and
concentrate the template DNA. The amount of template purified by our PCR product
recovery kit can be as much as 1/2 of the volume of the PCR reaction system.
- 2. The amount of template DNA: a very small amount of DNA can also be used as a template
for PCR, but to ensure the stability of the reaction, it is recommended to use target
sequences with more than 104 copies as a template for a 50μl system. Recommended
amount of template DNA.