一种适于PCR扩增的快速提取猪基因组DNA的碱裂解法

发布时间：2018-01-20 05:44

本文关键词： 猪基因组DNA 碱裂解法酚-氯仿法 PCR扩增　出处：《华中农业大学学报》2017年04期 　论文类型：期刊论文

【摘要】：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。
[Abstract]:The porcine genomic DNA was extracted from the longissimus dorsi muscle, spleen and ear tissue by alkaline lysis method and phenol-chloroform method, respectively. The genomic DNA was extracted by UV spectrophotometer and agarose gel electrophoresis. The yield and quality of DNA were detected by PCR amplification. The results showed that there was no significant difference in the yield of DNA extracted by the two methods. The target bands could be obtained by PCR amplification with TEF-1 gene specific primers. DNA extracted by alkaline lysis was diluted by 5 times. After 10 times as template, stable amplification products were obtained. The results showed that the simple, rapid and non-toxic alkaline lysis method was suitable for the extraction of DNA in animal qualitative PCR detection.
【作者单位】：河南牧业经济学院动物科技学院;华中农业大学农业动物遗传育种与繁殖教育部重点实验室;生猪健康养殖协同创新中心;华中农业大学实验动物中心;
【基金】：湖北省科技支撑计划项目(2014ABA033;2014ABA025)
【分类号】：Q78;S828
【正文快照】： 提取动物基因组DNA的方法有多种,其中经典的方法有酚抽提法、甲酰胺裂解法、异丙醇沉淀法、盐酸胍裂解玻璃棒缠绕法及酚-氯仿法等[1],这些方法都能获得高质量DNA;但是提取过程繁琐,技术、设备要求高、耗时长,且有的试剂具有一定的毒性。在经典的DNA提取方法上经过改进、优化出

【相似文献】