BloodTaq DNA Polymerase
BloodTaq DNA Polymerase is a novel DNA polymerase obtained through the deletion of an amino acid at the N-terminus of Taq DNA Polymerase and mutation modification. After modification, this product can withstand the inhibitors present in whole blood, and can directly amplify DNA in human and mouse whole blood samples without the need for prior genome extraction and purification. The 3 'end of the PCR product is A and can be directly used for T/A cloning.
Component specifications
BloodTaq DNA Polymerase (5U/ μ L) 5 × one hundred μ L
ten × BloodTaq PCR Buffer (with Mg2+) 5 × 1.8mL
Storage: -20 ℃
After multiple column purification, the purity was detected by SDS-PAGE to be over 99%;
No exogenous nuclease activity was detected;
PCR method for detecting non host residual DNA;
Effectively amplifying single copy genes in the human genome;
Store at room temperature for one week without significant changes in activity.
Before use, please invert the BloodTaq DNA Polymerase repeatedly until completely mixed.
Place the PCR thin-walled tube on ice and add the following reagents except for whole blood.
Component dosage and final concentration
BloodTaq DNA Polymerase (5U/ μ L) 1 μ L 5U
ten × BloodTaq PCR Buffer (with Mg2+) 5 μ L 1 ×
DNTP Mix (2.5mM each) 4 μ L 200 μ M each
Forward Primer (10 μ M) 2 μ L 0.4 μ M
Reverse Primer (10 μ M) 2 μ L 0.4 μ M
Whole blood ≤ 10%-
RNase Free water x μ L-
Total volume 50 μ L-
Attention:
Before adding whole blood, repeatedly suck and beat up and down to thoroughly mix various reagents.
DNA template: Whole blood can be treated with heparin sodium, Na EDTA, K-EDTA, or sodium citrate. It is usually recommended to have a whole blood content of 5-10%. It is not recommended to use high concentration blood. For templates with high GC content, add 10% DMSO.
Primers: Oligonucleotide primers typically contain 20-30 nucleotides in length, and preferably have a GC content of 40-60% and are evenly distributed within the primers. In conventional PCR reactions, the primer concentration should be between 0.1 and 1.0 at the final concentration μ M serves as a reference for setting the range.
ten × The BloodTaq PCR Buffer contains 30mM MgCl2.
Finally, add whole blood to the bottom of the tube.
PCR reaction conditions
Step Temperature Time Cycles
Pre denaturation at 95 ℃ for 5 minutes 1
Denaturation 95 ℃ for 30s 35-40
Annealing at 50-68 ℃ for 30 seconds
Extension 72 ℃ 250-500bp/min
Final extension 72 ℃ for 10 minutes
