Establishment of ISSR-PCR Reaction System and Screening of Effective Primers with ISSR Marker for Dracaena cambodiana

Abstract:

The influence factors of ISSR-PCR for Dracaena cambodiana Pierre ex Gagnep were studied by using single factor test and orthogonal design, and a set of ISSR-PCR reaction system was established. Sixteen ISSR primers were selected out, and their annealing temperatures were also optimized. Furthermore, the optimized reaction conditions were examined for its repeatability in D. cambodiana. The optimum ISSR-PCR reaction system with total 25 μL reaction solution contain 2.5 μL 10×PCR buffer, 3.0 mmol/L MgCl2, 300 μmol/L dNTPs, 3.0 U Taq polymerase, 0.2 μmol/L primers, and 20 ng DNA template, which can be used for analysis in genetic diversity and genetic structure of D. cambodianas.

Key words:Dracaena cambodiana; ISSR; Influence factor; Optimization system; Screening of primers