海洋底泥环境DNA提取与纯化方法比较研究

Comparative study of methods for extraction and purification of environmental DNA from marine sediment samples

为了探索有效的海洋底泥宏基因组DNA提取与纯化方法,分别采用CTAB结合SDS法、CTAB结合玻璃珠法、SDS结合蛋白酶K法和SoilMasterTM试剂盒法提取海洋底泥宏基因组DNA,用柱式腐殖酸清除试剂盒法与电洗脱法纯化宏基因组DNA,并对宏基因组DNA的产量与纯度进行评价.结果表明:SDS结合蛋白酶K法提取率最高,DNA片段完整;SoilMasterTM试剂盒法与CTAB结合SDS法次之;CTAB结合玻璃珠法有严重降解;电洗脱法可以得到高纯度、大于42kb宏基因组DNA片段.因此,SDS结合蛋白酶K法和电洗脱法提取纯化海洋近海底泥宏基因组DNA优于其他方法.

In order to seek an effective extraction and purification method of metagenomic DNA from marine sediment samples,the metagenomic DNA was extracted by CTAB plus SDS method,CTAB plus glass beads method,SDS plus proteinase K method and SoilMaster^TM DNA Extraction Kit method,respectively.The metagenomic DNA was purified by column humic clear kit method and electroelution.The quality and quantity of the extracted metagenomic DNA are compared.The highest extracted rate and integrity are obtained by SDS plus proteinase K method,next are SoilMaster^TM DNA Extraction Kit method and CTAB plus SDS method;CTAB plus glass beads method has severe degradation;and electroelution produces the purest metagenomic DNA and large fragments with more than 42kb.The research results show that SDS plus proteinase K method and electroelution are the optimum extraction and purification methods of metagenomic DNA from marine sediment samples compared with other methods.