一种提取动物基因组总DNA的野外样品保存方法

A Suitable Storage Method of Animal Sample for Extracting Genomic DNA in Field

为了确定一种方便的野外动物样品保存方法,以新鲜材料作对照,从-20℃冰箱、70%乙醇、含50mmol/L EDTA的70%乙醇、95%乙醇、液氮处理的高原鼠肌肉和肝脏组织中提取基因组总DNA。通过琼脂糖凝胶电泳和紫外分光光度计对提取的基因组总DNA质量进行检测。结果显示:相同处理的肝脏DNA产量大,肌肉组织提取的DNA质量好;各种保存方法提取的DNA降解程度依次为,-20℃冰箱、70%乙醇>含50mmol/L EDTA的70%乙醇、95%乙醇>液氮>新鲜。选择新鲜肌肉和95%酒精处理的肌肉样品提取的总DNA作模板,进行微卫星PCR扩增,均可获得清晰的电泳带。将该方法用于高原鼢鼠,进行线粒体12S rRNA、Cytb和D-loop区测序,结果显示该方法保存的样品与新鲜样品没有差别。因此,在野外用95%乙醇固定肌肉样品是一种可行的样品保存方法。

In order to select a suitable method of keeping animal samples in the field,genomic DNA was extracted from muscle and livers of plateau pika which were frozen in liquid nitrogen and-20℃,fixed with 70% or 95% ethanol,preserved in 50 mmol/L EDTA of 70% ethanol,and the fresh muscle and livers used as control samples.The extracted total DNA was examined with gelose gel electrophoresis and ultraviolet spectrophotometer.The results indicated that DNA could be got from all the samples,whereas the quality and quant...