Abstract:

Uniform design was applied to optimize SRAP(Sequence-related amplified polymorphism)-PCR(polymerase chain reaction) system of cynodon dactylon. Five main factors influenced on SRAP-PCR, including MgCl2, primer, dNTP, TaqDNA polymerase and DNA template were tested using uniform design U16(45 ) and U12(35 ). A 25 ul suitable SRAP-PCR system for cynodon dactylon was established. The SRAP-PCR amplification reaction system was consisted of 3.5 mmol/L MgCl2，0.20mmol/L dNTPs, 0.44umol/L primer,1.50U TaqDNA polymerase and 28g/L template. Annealing temperature was further researched and found that it has no significant influence on amplification results. By using this optimal system，the genomic DNA of other 10 samples of cynodon dactylon was amplified，and DNA bands with high polymorphism were clear. In conclusion, the optimization of SRAP-PCR amplification system can be used to study genetic relationship，systematic evolution and genetic diversity of cynodon dactylon.