| Description | This product is a premixed system composed of Es Taq DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers, with a concentration of 2 ×. Es Taq DNA Polymerase has excellent performance of high amplification efficiency and low mismatch rate. The unique MasterMix formula makes the entire This product is a premixed system composed of Es Taq DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers, with a concentration of 2 ×. Es Taq DNA Polymerase has excellent performance of high amplification efficiency and low mismatch rate. 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 does not contain dyes. After the PCR program is completed, an appropriate amount of sample buffer can be added as needed for electrophoresis operation. 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.E665629Component5ml25mlStorageE665629A2×Es Taq MasterMix5×1ml5×5ml-20℃. Avoid freeze/thaw cycle.E665629BddH₂O5×1ml5×5ml-20℃. Avoid freeze/thaw cycle.Notes: 2×Es Taq MasterMix contains Es Taq DNA Polymerase, 3mM MgCl2 and 400µM each dNTP.Quality control:After testing, there was no exogenous nuclease activity; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes from multiple genomes.Usage:The 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 system Reagent 50 µlReaction system Final concentration 2×Es Taq MasterMix 25 µl 1× Forward Primer，10 µM 2 µl 0.4 µM Reverse Primer，10 µM 2 µl 0.4 µM Template DNA <0.5 µg <0.5 µg/50 µl ddH2O up 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 conditions Step Temperature Time / Pre denaturation 95℃ 2 min / Denaturation 94℃ 30 s 25-35 cycles Anneal 55-65℃ 30 s 25-35 cycles Extend 72℃ 30 s 25-35 cycles Finally extended 72℃ 2 min / Attention: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 Es Taq 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... Read More | Inquire | Product Application：KNK437 has been used: as a heat shock factor 1 (HSF1) inhibitor to study its effects on the inhibition of viability and apoptosis activation in chemoresistant mice cells as an HSF1 inhibitor to study its effects on viability and apoptosis of colorectal cancer cells as a Product Application：KNK437 has been used: as a heat shock factor 1 (HSF1) inhibitor to study its effects on the inhibition of viability and apoptosis activation in chemoresistant mice cells as an HSF1 inhibitor to study its effects on viability and apoptosis of colorectal cancer cells as a heat shock protein 70 (HSP70) inhibitor to study its effects on glutamine-induced HSP70 and inflammatory mediator release... Read More | Inquire | Inquire |