毛干DNA的快速提取及其对PCR的影响

A Fast Hair Shaft DNA Extraction Method and Its Influence on PCR Amplification

[目的]建立毛干DNA快速稳定的提取扩增策略,使这一简便取材能在畜牧、野生动物保护等采样困难的领域得以普及应用。[方法]采用PCR缓冲液快速提取及2轮PCR逐级稀释扩增的策略提取扩增黄牛毛干DNA微卫星位点。[结果]该方法可从低至0.1mg(约1 cm长)的毛干中提取得到DNA,并成功扩增核DNA微卫星位点。48份黄牛牛毛样本均成功提得DNA。72份扩增样本,除4份样本扩增效果较差,其余样品均能正常扩增。[结论]该研究方法可快速稳定的提取扩增目标基因,为毛干在相关领域的普及应用奠定了基础。

[Objective] To establish a fast and stable hair shaft DNA extraction and amplification strategy,and make it to use in animal husbandry and wild animals protection.[Method] An improved PCR buffer fast extraction method and 2 rounds of PCR strategy were adopted to extract and amplify DNA locus of cattle hair shaft.[Result] The result showed that this strategy even can extract nuDNA （nuclear DNA） and amplify its target gcnes from 0.1 mg （about 1 cm） hair shaft.It successfully extracted DNA from all 48 hair shaft samples of cattle.In amplification stage,68 of 72 amplification samples have got its target genes.[Conclusion] This method can extract nuDNA from hair shaft and amplify its target genes fast and stable,which lays a foundation for application of hair shaft in relevant field.