Abstract: Objective　To establish a method of genome extraction for genotyping of germ cells of laboratory animals.Methods　The genomic phenol extraction method, column kit extraction method and magnetic bead kit extraction method were optimized and compared for the DNA extraction of sperm and embryos samples in terms of fluorescent PCR method.Results　1) All the three method above are effective for sperm DNA extraction. The column extraction kit optimized by adding DTT showes the highest genome extraction efficiency for sperm samples(P<0.05). 2) Column kit and magnetic bead kit could extract the embryo genome effectively, and the DNA magnetic bead kit had the highest yield when extracting embryos (P<0.05). Conclusion　The column extraction method optimized by adding dithiothreitol (DTT) is preferred when extracting sperm genome. When extracting the embryo genome, the DNA magnetic bead kit is preferred. All above provides a reference for genome extraction from frozen germ cells.

Key words: sperm, embryo, DNA extraction 

摘要： 目的　优化并比较3种基因组提取方法对冷冻生殖细胞的提取效率。方法　用苯酚抽提法、柱式试剂盒和 磁珠试剂盒经优化分别提取冷冻精子、胚胎样本基因组,通过荧光定量PCR比较以上3种方式提取效率。结果 1)3种方式均可提取精子基因组,通过加入二硫苏糖醇(DTT)优化的柱式提取法对精子样本的基因组提取效率最 高(P<0.05)。2)柱式试剂盒和磁珠试剂盒可有效提取胚胎基因组,在提取微量胚胎时,DNA磁珠试剂盒相比柱式 试剂盒得率较高(P<0.05)。结论　在提取精子基因组时,可优先选用加入二硫苏糖醇(DTT)前处理的柱式提取 法;在提取胚胎基因组时,可优先选用DNA磁珠试剂盒,该研究为冷冻生殖细胞基因组提取提供了参考。

关键词: 精子, 胚胎, 基因组提取