几种苎麻园土壤微生物总DNA提取方法比较

Comparison of Methods of DNA Extraction from Ramie Soil

实验设计对比了苎麻园土壤微生物总DNA6种提取方法。方法Ⅰ采用裂解酶、蛋白酶K、SDS与CTAB处理;方法Ⅱ结合利用CTAB、SDS、LDS和BME;方法Ⅲ综合采用玻璃珠、CTAB和SDS法;方法IV结合利用SDS-GITC-PEG;方法V利用尿素、SDS、LDS和BME综合处理;方法VI则利用试剂盒处理土壤。6种方法都可以提取到23kb左右的DNA片段,经过TENP和PVPP方法预处理的土壤样品,能有效去除腐殖酸等污染物,且提取得到的土壤总DNA完整性好、纯度高,不需要纯化就可用于PCR扩增。但不同方法取得的DNA的片段长度、产量和纯度存在明显差异。通过比较,改进并建立了可用于宏基因组构建的高纯度、大片段DNA的方法。

In this experiment,the effectives of six soil DNA extraction methods for isolation of the total microbial DNA from ramie plot were compared.Method I uses cleavage enzyme,protease K,SDS and CTAB;Method II uses CTAB,SDS,LDS,and BME;Method III uses glass beads,CTAB,and SDS;Method IV uses SDS-GITC-PEG;Method V uses Urea,SDS,LDS,and BME;Method VI uses kit on soil.The results show that the length of DNA is about 23 kb by all the six methods.The pretreatments treated by TENP and PVPP effectively remove contaminant...