SuperStar DNA Polymerase
This product is a Taq DNA Polymerase using the latest sealing technology, suitable for HotStart PCR. At temperatures below 70 ℃, the active center of the enzyme is completely blocked, effectively inhibiting polymerase activity and effectively inhibiting non-specific annealing of primers and non-specific amplification caused by primer dimers under low temperature conditions. After incubation at 95 ℃ for 5 minutes, some of the enzyme activity was released. As the thermal cycle continues, the remaining enzyme activity will be continuously released, which overcomes the disadvantage of conventional Taq enzymes being prone to plateau stage and greatly improves their amplification efficiency.
When PCR amplification is performed on this product, the 3 'end of the PCR product contains an "A" base, which can be directly used for T/A cloning. This product has the characteristics of fast amplification speed, good specificity, and good stability, and is mainly used for PCR and RT-PCR reactions that require a large amount of amplification products.
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.
Component 500U 2500U
SuperStar DNA Polymerase (5U/ μ L) 100 μ L 5 × one hundred μ L
ten × PCR Buffer 1.8mL 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;
Can effectively amplify single copy genes in the human genome.
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.
Preparation of PCR reaction system
Component dosage and final concentration
ten × PCR Buffer 5 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
SuperStar DNA Polymerase (5U/ μ L) 0.5 μ L 2.5U/50 μ L
DdH2O to 50 μ L
Attention:
The reaction solution can be prepared at room temperature, preferably by placing the reagent on ice. Primer concentration should be between 0.1 and 1.0 at the final concentration μ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the concentration of primers can be reduced to optimize the reaction system.
10% of this product × The PCR buffer contains 15mM magnesium ions.
Set up PC