一种直接用于PCR分析的风沙土微生物总DNA 提取方法 - Journal of .PDFVIP

第31卷第11期 东 北 大 学 学 报 ( 自 然 科 学 版 ) Vol31, No. 11 2 0 10 年 11 月 Journal of Northeastern U ni ersity( Natural Science) No . 2 0 1 0 PCR DNA 张 颖, 曹成有 ( , 110004) : , DNA SDS CTAB K DNA , PEG , SDS CTAB K DNA , DNA DNA 0586~ 1311g DNA 23 Kb PEG DNA , DNA 16S rDNA PCR SDS CTAB K PEG DNA DNA : ; ; DNA ; DNA ; PCR : Q 938. 1 : A : 1005-3026( 2010) 11- 1640-04 A Method for Total Microbial DNA Extraction Directly Meeting the Requirement of PCR Amplification from an Aeolian Sandy Soil ZH A N G Ying, CA O Cheng-y ou ( School of Sciences, Northeastern Uni ersity , Shenyang 110004, China. Correspondent : CAO Cheng-you, professor, E-mail: caochengyou @ 163. com) Abstract The soil microbial DNA w as directly extracted from aeolian sandy soil. The DNA crude extracts were obtained by the cell lysis ia the repeated freezing-thaw ing process of SDS, lysozyme, CTAB, proteinase K and liquid nitrogen in combination w ith the crude extracts purified by PEG ( polyethylene gly col) . In this w ay the large fragment DNA w as obtained from - 1 the lytic cells with the DNA yield increased, i. e. , 0586~ 1311 gg ( dry soil) . In addition, the fragment size of DNA w as o er 23 Kb, and PEG w as helpful to remo e humic acid in DNA effecti ely. The unpurified DNA could be directly used as templates for PCR amplification of 16S rDNA. The method mentioned abo e w as simple and appropriate for DNA extraction from the