底泥水体中适用于PCR的不同形态DNA的提取方法Methods of Extraction Different Gene Types of Sediments and Water for PCR Amplification
冯凌;罗义;
摘要(Abstract):
提出直接从环境河流底泥和表层水体中同时提取不同形态DNA(胞内、胞外、染色体和质粒DNA)的有效方法.利用该方法提取了海河典型区域底泥的胞外DNA和胞内DNA及水体中细菌染色体DNA和质粒DNA.结果表明,NaH2PO4提取的胞外DNA分子量大于1kb,提取率40%~72%,没有共提取胞内DNA.胞内DNA提取的分子量均大于23.1kb,细菌质粒DNA与染色体DNA同时分离.16SrDNA与sul2扩增验证提取方法可靠性.16SrDNA扩增结果显示所有胞外DNA呈阴性,胞内DNA呈阳性.质粒DNA和染色体DNA的16SrDNA扩增显示,染色体DNA结果显阳性,质粒DNA呈阴性,sul2的结果相反.
关键词(KeyWords): 胞外DNA;胞内DNA;染色体DNA;质粒DNA;提取方法
基金项目(Foundation): 国家自然科学基金(No.2077704;No.30870363);; 天津市自然科学基金(No.08JCYBJC02700)
作者(Author): 冯凌;罗义;
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DOI:
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