Establishment and optimization of SCoT-PCR system in Fragaria
Start Codon Targeted Polymorphism （SCOT） is a novel gene-targeted molecular marker tech- nique. In this experiment, a L25 （5^6） or thogonal design was used for the optimal SCoT-PCR reaction system of strawberry with 11 Fragaria species and a cultivar as materials. The optimal PCR reaction mixture of SCoT-PCR system for strawberry was 2.0 mmol·L^-1 Mg^2＋,0.3 mmol·L^-1 dNTPs,0.75 U Taq polymerase, 0.75 μmol·L^-1 primer and 40 ng template DNA in 20 μL mixtures. Furthermore, the annealing temperature of each primer was optimized; the methods of electrophoresis and strawberry DNA extracting were improved. 22 primers were selected which were suitable to the SCoT-PCR reactions. The optimal reaction system with high polymorphism, stability and repeatability was obtained.
国家科技期刊平台
NSTL
万方数据
维普
