一种适于PCR扩增的快速提取猪基因组DNA的碱裂解法

以大白猪背最长肌、脾脏、耳组织为材料,用碱裂解法和酚-氯仿法分别提取猪基因组DNA,用紫外分光光度计、琼脂糖凝胶电泳、PCR扩增检测DNA的产量和质量,结果发现2种方法提取的DNA产量没有明显差异。通过猪MC4R、TEF-1基因特异引物进行PCR扩增均能得到目的条带。碱裂解法提取的DNA原液稀释5倍、10倍后作为模板均获得了较为稳定的扩增产物。该结果表明,简单、快速、无毒的碱裂解法适用于动物定性PCR检测时DNA的提取。

A rapid alkaline lysis method of extracting pig genomic DNA for PCR amplification

Alkaline lysis and conventional phenol-chloroform extraction methods were adopted to extract genomic DNA from the longissimus dorsi muscle,spleen and ear tissues from Large White pigs.The quantity and quality of DNA were determined by ultraviolet spectrophotometer,agarose gel electrophoresis and PCR amplification.The results showed that there was no significant difference of DNA quantity between the two DNA extraction methods.The gene fragments of MC4R and TEF-1 could be obtained by PCR amplification using genomic DNA extracted by the two different extraction methods.Most importantly,the amplification results were relatively stable when using 5 and 10 times diluted DNA extracted by alkaline lysis method.Therefore,the simple,rapid and non-toxic alkaline lysis method can be widely used in extraction and determination of DNA for qualitative analysis in animals.