泰泽病原体基因组DNA提取方法的建立

Establishment of Method of Extracting Genomic DNA from Tyzzer Organism

目的 提取泰泽病原体基因组DNA ,为建立该菌基因组文库奠定基础。方法 使用密度梯度离心结合酶解消化方法、酶解消化方法、本研究建立方法即过滤盐析离心法 ,从感染肝脏组织纯化泰泽病原体 ,并比较三种方法纯化泰泽病原体效果 ;采用氯化苄法、试剂盒、酚法提取泰泽病原体基因组DNA ,并比较三种方法提取基因组DNA质量 ;鉴定酚法提取泰泽病原体基因组DNA特异性。结果 使用过滤盐析离心法从感染肝脏组织纯化泰泽病原体 ,采用酚法提取其基因组DNA ,所获得的基因组DNA特异性好、纯度高、DNA片段长度大于 5 0kb ,且均一性好 ,无降解。结论 本研究首次成功提取泰泽病原体基因组DNA 。

Objective To construct DNA library ,genomic DNA was extracted from Tyzzer organism. Methods Three methods (density gradient centrifugation combining enzymatic digestion method, enzymatic digestion method, and filtration salting-out combining centrifugation method) were used to purify Tyzzer organism from infectious livers, and results of purification were compared. Genomic DNA was extracted from Tyzzer organism by three methods (benzyl chloride-method, kit and phenol method), character of which was analyzed. Results Tyzzer organism was purified effectively by filtration salting-out combining centrifugation method. Genomic DNA extracted by phenol method was high quality and specificity.Conclusion Genomic DNA was first extracted from Tyzzer organism successfully, which can apply to many experiments of molecular biology.