Xu Xiao-jiao, Cao Cheng-you, Cui Zhen-bo, Yang Min
College of Sciences, Northeastern University, Shenyang, China.
Ying Yong Sheng Tai Xue Bao. 2010 May;21(5):1327-33.
In order to select and establish an appropriate method for extracting and purifying the microbial total DNA from an aeolian sandy soil, six extraction methods (five direct methods and one indirect method) and two purification methods were examined, with the quantity and quality of extracted and purified total DNA compared. All the six extraction methods could extract the total DNA with a length of approximately 23 kb, among which, the improved SDS high salt extraction method (using 40% PEG8000 and 4 mol x L(-1) NaCl to precipitate DNA) was the best. This method could have a yield slightly less than that obtained by using kits, and the extracted DNA had the highest purity after purification, being available in 16S rDNA PCR amplification. Among the purification methods, the effect of agarose gel electrophoresis plus minicolumn was satisfactory, with most of the purified total DNA being able to be PCR-amplified and meet the requirements of the purity of DNA in the follow-up molecular operations.
为了选择并建立一种从风沙土中提取和纯化微生物总DNA的合适方法，研究了六种提取方法（五种直接法和一种间接法）和两种纯化方法，并比较了提取和纯化后总DNA的数量和质量。六种提取方法均能提取出长度约为23 kb的总DNA，其中改良SDS高盐提取法（使用40% PEG8000和4 mol·L⁻¹ NaCl沉淀DNA）效果最佳。该方法的产量略低于试剂盒提取法，纯化后提取的DNA纯度最高，可用于16S rDNA PCR扩增。在纯化方法中，琼脂糖凝胶电泳加微型柱的效果令人满意，大部分纯化后的总DNA能够进行PCR扩增，并满足后续分子操作中DNA纯度的要求。
