浙贝母根际土壤总DNA提取和纯化方法的比较

Comparison of Rhizospheric Soil DNA Isolation and Purification Methods from the Root of Fritillaria thunbergii Miq

摘要:

本研究通过对6种土壤总DNA提取方法(CTAB-SDS-冻融法、玻璃珠-SDS-酚氯仿抽提法、玻璃珠-聚乙烯聚吡咯烷酮-SDS法、玻璃珠-聚乙烯聚吡咯烷酮-溶菌酶法、玻璃珠-聚乙烯聚吡咯烷酮-冻融法和UltraClean试剂盒法)和4种纯化方法(改进琼脂糖凝胶电泳法、2％聚乙烯吡咯烷酮-琼脂糖凝胶电泳法、PVPP层析柱法和低熔点琼脂糖电泳法)的比较, 证明利用20 mmol/L EDTA (pH 7.5)预处理土壤后, 利用CTAB-SDS-冻融法提取土壤总DNA并经改进琼脂糖凝胶电泳法纯化获得的浙贝母根际土壤总DNA的得率和纯度相对较高, 达44.00 μg/g±2.65 μg/g土壤, 可用于后续基于16S rDNA分析基础上的土壤微生物分子生态学的分析工作。

Abstract:

Six DNA extraction methods and four DNA purification methods were compared and analyzed in this study to get higher quality DNA from the rhizospheric soil of Fritillaria thunbergii Miq. Results showed that higher purity DNA were harvested by pretreating the soil with 20 mmol/L EDTA (pH 7.5), then isolating soil DNA with CTAB-SDS-frozen-thawing, and further purified by agarose method. The recovery rate of this soil DNA was about 44.00 μg/g ± 2.65 μg/g soil, and they were qualified for the microbial diversity analysis in the rhizospheric soil of F. thunbergii Miq based on the 16S rDNA sequence.