HiFi PCR Mix for NGS (C09-1541-511)

Product content:Component:H665865-1ml:H665865-5ml:2xHiFi PCR Mix:1 ml:5x1 ml:ddH₂O:1 ml:5x1 ml:Product Introduction:HiFi PCR Mix for NGS is a premixed system consisting of hot starter enzyme, PCR Buffer, dNTPs, Mg2+, and PCR stabilizers and enhancers, etc. It is characterized by high fidelity, high elongation, and low preference, and has a balanced amplification efficiency for complex DNA templates (e.g., high GC-content templates), and it is especially suitable for multiplexed PCR in the thousand-two library building .The high efficiency hot starter enzyme contained in this product has no polymerase activity at room temperature, which effectively avoids non-specific amplification caused by non-specific binding of primers and templates or primer dimerization under room temperature conditions. The unique combination of buffer system and hot starter enzyme significantly improves the amplification efficiency of PCR with wider amplification range. This product effectively enhances the amplification efficiency of high GC or high AT regions in the genome, reduces amplification preference and thus improves sequencing coverage.matters needing attention:This product is not suitable for thousands of primers with modifications.Before use, please mix gently by turning up and down, avoid foaming as much as possible, and use after brief centrifugation. Avoid repeated freezing and thawing, which may deteriorate the performance of the product. If frequent use is required within a short period of time, store at 2-8°C.Usage:The following examples are conventional PCR reaction systems and reaction conditions, which should be improved and optimized according to the template, primer structure and target fragment size in actual operation.PCR reaction system:Reagents:50 μl Reaction system:2xHiFi PCR Mix:25 µI:Primer Pool:0.1-0.3 µM:DNA or cfDNA:5 ng-100 ng:ddH2O:up to 50 µl:Note: Please use the final concentration of 0.1-0.3µM as a reference for setting the range of primer concentration.2. PCR reaction conditions:Attention:In general, the annealing temperature was 5°C lower than the melting temperature Tm of the amplification primers, and when the desired amplification efficiency could not be obtained, the annealing temperature was lowered appropriately; when non-specific reactions occurred, the annealing temperature was increased, and the reaction conditions were optimized.2) The number of cycles can be set according to the downstream application of the amplification product. If the number of cycles is too low, the amplification is insufficient; if the number of cycles is too high, the chance of mismatch will increase and the non-specific background will be serious. Therefore, the number of cycles should be minimized under the premise of ensuring the product yield.