4种提取口腔产黑色素细菌DNA方法的比较  

Four methods for extraction and purification of DNA from Prevotella intermedia

作　　者：都佳寅[1] 王宁[2] 闫卉[1] 

机构地区：[1]天津市南开医院口腔科,天津300100 [2]天津中医药大学第二附属医院口腔科,天津300150

出　　处：《中国现代医学杂志》2015年第11期17-20,共4页China Journal of Modern Medicine

摘　　要：目的比较4种提取口腔产黑色素细菌中间普氏菌DNA的方法,即酚-氯仿法、溴化十六烷基三甲铵(CTAB)提取法、Chelex-100提取法及试剂盒。方法分别记录OD260、OD230、OD280及OD340值;聚合酶链反应(PCR)电泳,检测提取DNA的浓度、纯度及对PCR反映体系的影响。结果试剂盒及CTAB法去除黑色素比较差异无统计学意义(P>0.05),两者与酚-氯仿及Chelex-100法比较差异有统计学意义(P<0.05);CTAB法提取DNA的浓度及纯度的效果要优于试剂盒,差异有统计学意义(P<0.05),PCR电泳条带清晰,无拖带。结论 CTAB法可有效去除黑色素及其他杂质,同时获得较高浓度的DNA样品;黑色素并不溶于氯仿/异戊醇混合液,对PCR反应体系有很大影响。[Objective] To compare four DNA extraction methods and to select a more appropriate method for elimination of melanin created by Prevotella intermedia. [Methods] Phenol chloroform method, CTAB method, Chelex-100 method and a commercial kit were used to extract nuclear DNA of Prevotella intermedia with the same concentration. The efficiency of eliminating melanin was assessed by OD260, OD230, OD280, OD340, PCR assay and eleetrophoretic results. [Results] The efficiency of eliminating melanin by CTAB method was not significantly different from that by the commercial kit （P 〉 0.05）. The efficiency of eliminating melanin by CTAB method and the commercial kit was significantly different from that by phenol chloroform method and Chelex-100 method （P 〈 0.05）. The concentration and purity of DNA extracted by CTAB method were higher than those by the commercial kit （P〈 0.05） with clear PCR eleetrophoresis bands. [Conclusion] Melanin can significantly influence phenol chloroform method, Chelex-100 method and commercial kit. CTAB method can efficiently eliminate melanin and other foreign substances, and provide high-concentration DNA.

关 键 词：DNA提取 黑色素 聚合酶链反应 电泳 OD值 

分 类 号：Q78[生物学—分子生物学]