作　　者： ; ; ; ; ;

机构地区： 广东海洋大学食品科技学院

出　　处： 《生物技术通报》 2013年第12期119-122,共4页

摘　　要： 为获得高质量南美白对虾肠道微生物宏基因组模板,采用传统CTAB法、试剂盒和仪器法提取南美白对虾(Penaeus Vanmamei,P.vannamei)肠道微生物宏基因组,通过琼脂糖凝胶电泳、紫外分光光度法和V3区16S rDNA扩增检测所得基因组的完整性、得率、纯度和目的片段质量。结果表明3种方法提取得到的DNA结构均较完整,但得率和纯度差别较大,传统CTAB法提取的DNA得率和纯度均试剂盒和仪器法高,试剂盒和仪器法受糖和盐类污染较高,3种方法提取的宏基因组均可特异扩增出V3区保守片段。综合比较得知传统CTAB法提取P.Vanmamei肠道微生物基因组在完整性、得率和纯度方面优于其他两种方法,可为后续分子生物学分析提供良好基因组模板。 In order to obtain the high quality total genomic DNA template from Penaeus vannamei intestinal microbiota,the DNA extraction methods consisted of traditional CTAB,bio-kit,and apparatus were compared with in terms of integrity,yield,purity and target fragment quality by agarose gel electrophoresis and spectrophotometry detection. The results showed that total genome DNA could be well extracted with any of the three methods. The yield and purity,however,were significantly different. The traditional CTAB extraction had better performance than kit and apparatus methods,of which high pollution of carbohydrates and salts was observed,but 16S rDNA fragment in the conserved region of V3 could be specifically amplified with any method. The conclusion can be drawn that CTAB can satisfy the follow-up molecular biology experiments by offering high quality genome DNA template in integrity,yield,and purity.

关 键 词： 南美白对虾 肠道微生物 宏基因组提取

领　　域： [生物学]