Discussion on Main Influencing Factors of PCR Experiment

This article explored the various influencing factors of PCR experiment to find appropriate ratio of each factor concentration.The result of PCR is affected mainly by template DNA,Mg2+,Taq DNA polymerase,primer,4dntps mixture and buffer.Wheat mianyang 19 with 5+10 HMW-GS gene was taken as a material to extract the DNA.Six experiments on five conditions were designed to study the effects on PCR.The results show that it can not gain peculiar band if absent any factor in the reaction system,any factor is not the more the better.The fitful volume is important to enhance the result of experiment.The article suggests the optimalized PCR system with total volume 25 μL including 32 ng/μL template DNA,2 mmol/L Mg2+,0.3 mmol/L 4dNTPs,1.2 U/25 μL Taq DNA polymerse,0.8 μmol/μL primer,and 1×buffer.