Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2010, Vol. 26 ›› Issue (23): 48-52.

Special Issue: 生物技术 园艺

• 23 • Previous Articles     Next Articles

Comparison of Genomic DNA Extraction Method and Optimization of ISSR-PCR Amplification for Grapefruit

  

  • Received:2010-09-03 Revised:2010-09-29 Online:2010-12-05 Published:2010-12-05

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.