2×BalbMulti PCR Mix
BalbMulti PCR Mix is a premixed system composed of BaldStar DNA Polymerase, Mg2+, dNTPs, as well as PCR stabilizers and enhancers. The use of this product does not require the optimization process of complex PCR reaction conditions, and multiple PCR reactions can be easily carried out with simple condition exploration. Suitable for various types of multiple PCR reactions, such as microsatellite analysis, genotyping, and SNP detection.
The BaldStar DNA Polymerase contained in this product is a chemically modified hot start enzyme that can effectively reduce non-specific amplification caused by primer mismatch in the early stages of PCR reactions. The activation of the enzyme requires incubation at 95 ℃ for 10 minutes. This enzyme, in combination with PCR enhancers that can enhance reaction specificity and a unique buffer system, allows all primers in the reaction system to be effectively extended without additional optimization. This product also includes GC Enhancer, which helps to achieve efficient amplification of "difficult" templates (such as templates with high GC content).
Component specifications
two × BalbMulti PCR Mix 5 × 1mL
DdH2O 5 × 1mL
Storage: -20 ℃, avoid repeated freeze-thaw.
After testing, there was no exogenous nuclease activity;
PCR method for detecting non host residual DNA;
Storage at 2-8 ℃ for three months showed no significant change in activity.
The following examples are the conventional PCR reaction system and reaction conditions. In practical operation, corresponding improvements and optimizations should be made based on different templates, primer structures, and target fragment sizes.
PCR reaction system
Component dosage and final concentration
two × BalbMulti PCR Mix 25 μ L 1 ×
Primer Mix, 10 μ M each 1 μ L 0.2 μ M
Template DNA in moderation
DdH2O to 50 μ L
Attention: When designing primers, try to minimize the difference in Tm between each primer, and try to control the difference within 5 ℃. Please use the final concentration of 0.05-0.2 for each primer 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 amplification occurs, the primer concentration can be reduced to optimize the reaction system.
PCR reaction conditions
Step Temperature Time Cycles
Pre denaturation at 95 ℃ for 10min 1
Denaturation at 95 ℃ for 30s 30-40
Annealing at 55~65 ℃ for 30 seconds
Extension 72 ℃ 1kb/min
Final extension 72 ℃ for 5 minutes 1
Attention:
In general experiments, the annealing temperature is 5 ℃ lower than the melting temperature Tm of the amplification primer, and when ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When a non-specific reaction occurs, increase the annealing temperature to optimize the reaction conditions.
The extension time should be set based on the size of the amplified fragment, and the amplification efficiency of BaldStar DNA Polymerase contained in this product is 1-2 kb/min.
According to the amplification products
