磁铁矿尾矿宏基因组DNA提取方法的初步研究

Preliminary Study on Extraction Method of Metagenomic DNA from Tailings of Magnetite

[目的]寻找一种有效的磁铁矿尾矿宏基因组DNA的提取方法。[方法]分别采用直接提取法和SDS高-盐法提取邯邢矿区磁铁矿尾矿宏基因组DNA,比较2种方法的提取效果。[结果]直接提取法的提取效果较差,基本上没有提出DNA,SDS高-盐法的提取效果较好,DNA得率较高;23 S rDNA特异序列扩增结果显示,所提DNA能够满足PCR扩增的需要。[结论]所取尾矿样品中含有大量的金属离子,这些金属离子不但影响溶菌酶的活性,而且影响后续PCR扩增效率,所以提取DNA前需对样品进行高浓度TE处理。

[Objective]The aim was to seek one effective extraction method of metagenomic DNA from tailings of magnetite.[Method]The metagenomic DNA from tailings of magnetite in Hanxing mining area was extracted by direct extraction method and SDS-high salt extraction method.The extraction effects of 2 kinds of method were compared.[Result]The extraction effect of direct extraction method was worse,there was almost no DNA being extracted.The extraction effect of SDS-high salt extraction method was better,and the yield rate of DNA was higher.The results of 23 S rDNA specific sequences amplification showed that the extracted DNA could meet the requirements of PCR amplification.[Conclusion]The tested tailing samples contained large amounts of metal ion,and the metal ions not only had effect on activity of lysozyme,but also had effect on further PCR amplification efficiency.So the tailing samples should be treated by TE with high concn.