BaldStar Best DNA Polymerase
This product is a heat activated high fidelity polymerase that has undergone special treatment. This polymerase has 5 '-3' DNA polymerase activity, 5 '-3' exonuclease activity, and 3 '-5' exonuclease activity. Under ordinary PCR conditions, compared with BaldStar Taq DNA Polymerase, it has excellent performance of high amplification efficiency and low mismatch rate. Chemical modification causes the enzyme to have no polymerase activity at room temperature, effectively avoiding non-specific amplification caused by non-specific binding of primers and templates or primer dimers at room temperature. The activation of the enzyme must be incubated at 95 ℃ for 10 minutes, which can be integrated into existing PCR thermal cycling programs. The optimized buffer system maximizes the effectiveness of the enzyme, achieving high fidelity, specificity, amplification efficiency, and sensitivity for the target fragment. The PCR product amplified using this product has an "A" base attached to the 3 'end, which can be directly used for T/A cloning. This product is suitable for conventional PCR, RT-PCR, and multiplex PCR, especially for PCR with high requirements for specificity, fidelity, and amplification efficiency. 5 in this product × BaldStar PCR Buffer contains 8.5mM magnesium ions.
Component 500U 2500U
BaldStar Best DNA Polymerase (5U/ μ L) 100 μ L 5 × one hundred μ L
five × BaldStar PCR Buffer 1.9mL 5 × 1.9mL
Storage: -20 ℃, avoid repeated freeze-thaw.
Using activated salmon sperm DNA as a template/primer, the amount of enzyme required to incorporate 10 nmol of deoxynucleotides into acidic insoluble substances is defined as 1 active unit (U) at 74 ℃ for 30 minutes.
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 month without significant changes in activity.
The following example is a PCR reaction system and reaction conditions for amplifying a 1kb fragment using human genome DNA as a template. In practical operation, corresponding improvements and optimizations should be made based on the template, primer structure, and target fragment size.
PCR reaction system
Component dosage and final concentration
five × BaldStar PCR Buffer 10 μ L 1 ×
DNTP Mix, 10mM each 1 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
Template DNA ＜ 0.5 μ G ＜ 0.5 μ G/50 μ L
BaldStar Best DNA Polymerase (5U/ μ L) 0.5 μ L 2.5U/50 μ
