Establishment and optimization of ISSR reaction system for Agropyron cristatum (L.) Gaertn.

It is necessary to establish and optimize the ISSR-PCR reaction system for genetic diversity study of Agropyron cristatum(L.) Gaertn.Results show that the optimum concentration of seven reactants in 20 μL reaction mixture were as follows: 40 ng genomic DNA,0.2 mmol·L-1 dNTP,0.5 μmol·L-1 ISSR primer,1 U Taq DNA polymerase,2 μL 10× PCR buffer,2.5 mmol·L-1 MgCl2.The suitable annealing temperature of primer 815 was 50℃.This reaction system would provide the basis for the diversity analysis of A.cristatum with ISSR markers.