Abstract:

An efficient genomic DNA extraction method useful for ISSR-PCR of grapefruit was selected by comparing four DNA extraction methods. Meanwhile, the factors influencing ISSR-PCR for grapefruit were optimized by orthogonal design. The result showed that improved SDS method was suitable for ISSR-PCR amplification. The average amplified bands were 2.7 which were detected in 2% agarose. The optimum amplification condition of ISSR-PCR was including 5 ng DNA template, 0.2 mmol/L dNTPs, 0.2 μmol/L primer, 0.3 U Taq DNA polymerase and 2×buffer (Mg2+) in 20 μL reaction volumes.