A method suitable for DNA extraction from humus-rich soil
A method suitable for DNA extraction from humus-rich soil
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一种适合从富含腐殖质的土壤中提取DNA的方法
DOI：
10.1007/s10529-014-1591-5
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发表时间：
2014-11-01
影响因子：
2.7
通讯作者：
Yao, Shuo
中科院分区：
文献类型：
--
作者：
Miao, Tianjin;Gao, Song;Yao, Shuo
提出了一种快速、简便的土壤DNA提取方法。在EDTA、SDS、苯酚、氯仿和异戊醇（3-甲基-1-丁醇）存在下，通过直接细胞裂解，然后用2-丙醇沉淀来提取土壤DNA。用改良的DNA纯化试剂盒和DNA凝胶提取试剂盒对提取的DNA进行纯化。利用该方法，从富含腐殖质的暗棕色森林土壤中提取的DNA不含腐殖物质，因此可用于高效的PCR扩增和限制性酶切。相比之下，传统的CTAB法提取的样品DNA的得率和纯度较低，且无法从同一土壤样品中提取出常用的商业化土壤DNA提取试剂盒。此外，该方法省时、方便，特别是为富含腐殖质的土壤中DNA的提取提供了一种有效的选择。
A rapid and convenient method for extracting DNA from soil is presented. Soil DNA is extracted by direct cell lysis in the presence of EDTA, SDS, phenol, chloroform and isoamyl alcohol (3-methyl-1-butanol) followed by precipitation with 2-propanol. The extracted DNA is purified by modified DNA purification kit and DNA gel extraction kit. With this method, DNA extracted from humus-rich dark brown forest soil was free from humic substances and, therefore, could be used for efficient PCR amplification and restriction digestion. In contrast, DNA sample extracted with the traditional CTAB-based method had lower yield and purity, and no DNA could be extracted from the same soil sample with a commonly-used commercial soil DNA isolation kit. In addition, this method is time-saving and convenient, providing an efficient choice especially for DNA extraction from humus-rich soils.