一种快速提取南瓜大群体基因组DNA的方法

摘要

基因型鉴定是作物分子标记辅助育种和基因功能遗传分析中十分重要的环节,此项技术的应用正日渐频繁,因而迫切需要一种高通量筛选的方法.本试验以南瓜中筛选到的互补型SSR引物为验证手段,在种子阶段比较了钻孔法和萌芽法采用碱煮法提取DNA的效果,并且在成熟植株阶段验证了各组织部位采用碱煮法提取DNA的效果.结果表明:使用萌芽的南瓜种子根尖以碱煮法提取DNA是可用于种子纯度测定的快速方法;成熟植株各部位采用碱煮法提取DNA均可用于植株基因型筛选.%Genotype identification is an important link in molecular-marker-assisted breeding and genetic analysis of gene function.Since this method is used more and more frequently,researchers need a high-through-put technique.This experiment took a pair of SSR primers of complementary type as the method,and compared the effect of extracting DNA from cotyledon and root during seedling stage.The effect of extracting DNA from each tissue of the plant by NaOH-boiling method during maturing stage was also verified.The results indicated that extracting DNA from root with NaOH-boiling method was a rapid way to measure seed purity.Therefore,extracting DNA with NaOH-boiling method from all tissues of the matured plant could be used for screening plant genotype.