板栗SSR和RAPD技术体系的建立Establishment of SSR and RAPD technique system in Castancea mollissima
王同坤;董超华;马艳;齐永顺;张京政;柏素花;
摘要(Abstract)：
主要从板栗DNA提取、PCR条件的优化等环节对SSR和RAPD的反应条件进行了优化。结果表明,采用PVP和β-巯基乙醇联合除酚,用多次洗涤和高盐相结合进行除糖提取高质量板栗基因组DNA,所获得的DNA完整,无降解,分子量在23kb以上,可扩增、可酶切,D260nm/D230nm在2.0以上,D260nm/D280nm为1.8～2.0,产率为40～300μgDNA/g叶组织,完全满足SSR、RAPD、AFLP等分子标记技术分析的需要。采用单因素对PCR反应体系中各个影响因素进行优化,结果表明对PCR结果影响最为明显的是退火温度和Mg2+浓度的变化。对退火温度和Mg2+浓度进行优化,确定RAPD的PCR反应体系Mg2+浓度为2.0mmol/LMgCl2,退火温度38℃;SSR反应体系Mg2+浓度为2.0mmol/L,退火温度56℃。建立了一套完善的适用于板栗的SSR和RAPD分析的稳定技术体系。
关键词(KeyWords)： 板栗;SSR;RAPD
基金项目(Foundation): 河北省自然科学基金资助项目(C2004000404)。
作者(Author): 王同坤;董超华;马艳;齐永顺;张京政;柏素花;
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DOI:
参考文献(References)：
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