Establishment and Optimization of SRAP-PCR System and Primer Screening for Vaccinium ssp
The research was conducted for further research of Vaccinium ssp in molecular genetics and marker assistant select breeding by marker technology. An optimized PCR system of SRAP marker (Sequence-Related Amplified Polymorphism) for Vaccinium ssp and the primer combination of screening were established- The genomic DNA was extracted from the young leaves by modified CTAB method, and three factors affecting SRAP-PCR were analyzed by single-factor test including the concentration of template DNA, primer concentration and annealing temperature. The optimized SRAP-PCR system (total 20 μL) was established including 0.40 μmol/L primer concentration, 120 ng template DNA, 10 μL 2×Taq PCR Master Mix, and deionized water for making up the remaining volume. Optimal annealing temperature of PCR amplification program was 50 ℃. 15 primer combinations with distinct and rich polymorphism were screened out from 80 primer combinations of SRAP.
NETL
NSTL
万方数据
维普
