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本试验对青花菜基因组DNA的SRAP-PCR体系中主要影响因子Mg2+、dNTPs和引物浓度进行了优化。结果表明,反应体系中适宜浓度为Mg2+1.5-3.0mmol/L,dNTPs0.4mmol/L,引物0.25-0.50μmol/L(模板DNA约20ng,16μL反应体系)。运用优化的反应体系,对20个青花菜品种进行分子鉴定,从10个引物组合中筛选到7个多态性引物组合,获得60个多态性位点,平均每个引物组合在供试品种中产生8.6个多态性位点,鉴别品种数4.3个。双引物组合me1/em6与me2/em9可以区分所有供试材料。
In this study, the main factors Mg2 +, dNTPs and primer concentration in SRAP-PCR system of broccoli genomic DNA were optimized. The results showed that the suitable concentration of Mg2 + 1.5-3.0mmol / L, dNTPs 0.4mmol / L, primer 0.25-0.50μmol / L (template DNA about 20ng, 16μL reaction system). The optimized reaction system was used to identify 20 cultivars of broccoli. Seven polymorphic primer combinations were screened from 10 primer combinations to obtain 60 polymorphic loci. The average number of each primer combination in the tested varieties 8.6 polymorphic loci were generated, identifying 4.3 varieties. The double primer combination me1 / em6 and me2 / em9 can distinguish all the tested materials.