| In order to optimize SRAP-PCR reaction system for Renanthera Loureiro, the U20(55) uniform experimental design was applied to optimize concentration of DNA template, Mg2+, primers, Taq DNA polymerase and dNTPs. Then the system was verified with 10 R. germplasm genome DNA as template and three pairs of SRAP primers. The results showed that the extraction of genomic DNA was of high purity and had good integrity; The bands of 20 treatments amplifing with Me6/Em6 primers, were clear, polymorphic processing of rich; Further screening using 2 DNA template for getting, the best system was the total volume of 20 μL reaction solution containing 60 ng genomic templates DNA, 2.0 mmol/L Mg2+, 1.0 μmol/L Primers, 1.2 U Taq DNA polymerase, 0.20 mmol/L dNTP. The bands obtained in the verification experiment were clear and highly polymorphic. |
