酿酒葡萄DNA的提取方法

Optimization of genomic DNA extraction from grapes

采用琼脂糖凝胶电泳、核酸蛋白检测和聚合酶链反应扩增(polymerase chain reaction,PCR)检测DNA的浓度和纯度,对改良十六烷基三甲基溴化铵(hexadecyl trimethyl ammonium bromide,CTAB)法与2种商业试剂盒提取4种不同品种酿酒葡萄DNA的方法进行了对比分析,结果表明改良的CTAB法提取DNA完整性较好,基因组DNA样品的OD260/OD280值均在1. 8左右,4个酿酒葡萄均能扩增出PCR条带,且条带清晰,无拖尾现象。说明改良的CTAB法提取的酿酒葡萄基因组DNA浓度和纯度均较高,可以进行PCR扩增、电泳检测、遗传多样性分析以及基因图谱构建等下游分子生物学研究试验。

An improved CTAB method was developed for efficient extraction of genomic DNA from four different types of grapes. The quality of DNA was detected by agarose gel electrophoresis and nucleic acid/protein analyzer.The ratio of OD260/OD280 of the extracted DNA reached 1. 8. The DNA on agarose electrophoresis was clear and suitable as template for specific gene amplification. The quality of extracted DNA by the improved CTAB method reached the requirement of subsequent molecular biological researches,such as PCR amplification,restriction pattern analysis,genetic diversity analysis and genetic map construction.