Description
This product is a premixed system composed of Taq Plus DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers, with a concentration of 2 ×. Compared with Taq DNA Polymerase, Taq Plus DNA Polymerase has higher amplification efficiency and better fidelity. The unique MasterMix formula makes the entire reaction system very stable, with over 98% of PCR amplification successful at once. At the same time, complex templates can also be effectively amplified, and human error and contamination can be minimized to the greatest extent. This product has been added with a dye (blue), and can be directly subjected to electrophoresis detection after the reaction is completed. Most PCR products obtained from amplification have an "A" base attached to the 3 'end, making them suitable for direct use in T/A cloning. Mainly suitable for routine PCR reactions and gene cloning experiments that require high fidelity.Quality controlAfter testing, there was no exogenous nuclease activity; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes from multiple genomes.UsageThe following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment using human genomic DNA as a template. In practical operation, corresponding improvements and optimizations should be made based on the template, primer structure, and target fragment size.1. PCR reaction systemreagent50 µl Reaction systemfinal concentration2×Taq Plus MasterMix（Dye）25 µl1×Forward Primer，10 µM2 µl0.4 µMReverse Primer，10 µM2 µl0.4 µMTemplate DNA<0.5 µg<0.5 µg/50 µddH2Oup to 50 µl/Attention: The primer concentration should be between 0.1 and 1.0 as 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 primer concentration can be reduced to optimize the reaction system.2. PCR reaction conditionssteptemperaturetime Pre denaturation94°C2 min Degeneration regression94°C30 s25-35 cyclesFire extension55-65℃30 s25-35 cyclesFinal extension72°C30 s25-35 cyclesextend72°C2 minAttention:1) In general experiments, if the annealing temperature is 5 ℃ lower than the melting temperature Tm of the amplification primer, and the ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When non-specific reactions occur, increase the annealing temperature to optimize the reaction conditions.2) The extension time should be set according to the size of the amplified fragment, and the amplification efficiency of Taq Plus DNA Polymerase is 2 kb/min.3) The number of cycles can be set based on the downstream application of the amplification product. If the number of cycles is too small, the amplification amount is insufficient; If there are too many cycles, the probability of mismatches will increase, and non-specific backgrounds will be severe. So, while ensuring product yield, the number of cycles should be minimized as much as possible