Optimization and application of ISSR-PCR reaction system in pear cultivars

The influencing factors in ISSR reaction system were studied by using genomic DNA of different pear cultivars. ISSR-PCR optimized reaction systems were eatablished for pear cultivars. The total volume of 25 滋L included 2.5 滋L10伊buffer, 2.5 mmol/L MgCl2, 0.2 mmol/L dNTPs, 0.25 滋mol/L primer, 10~60 ng template DNA and 0.75 U unit of DNA polymerase. 22 different pear cultivars were used to check the stabilization of the optimized reaction system. The results showed that the optimal reaction system of ISSR-PCR of pear was stable, repeatable and effective .