An economic and efficient method for further purification of crude DNA extracted from forest soils  

一种经济有效的森林土壤粗品DNA纯化方法(英文)

作　　者：韩国民[1] 宋福强[2] 倪武[1] 何沙娥[3] 张智俊[3] 田兴军[1] 

机构地区：[1]南京大学生命科学学院,南京210093 [2]黑龙江大学生命科学学院,哈尔滨150000 [3]浙江林学院浙江省现代森林培养重点实验室,临安311300

出　　处：《Journal of Forestry Research》2010年第2期246-250,I0008,共6页林业研究（英文版）

基　　金：supported by Project of National Science Foundation of China (30870419, 40971151);the National Basic Research Program of China (2008CB418004);the Open Research Foundation of Key Laboratory for Modern Sylvicultural Technology of Zhejiang Province (200605), Zhejiang Forestry College,China

摘　　要：To obtain pure DNA directly from some complex forest soils are still very difficulty at present,though many methods even commercial kits have been attempted.This paper reports an economic and efficient method for further purifying crude DNA extracted from forest soils with two steps.First,the crude DNA was dissolved using the extraction buffer,which removed the debris by chloroform-isoamyl alcohol,and then reprecipitated the DNA by isopropanol;second,the recovered DNA was further purified with silica spin column.Results show that 82-91% of the humic acids was removed by step one.The remaining humic acids could be completely effaced through the second step.The recovered DNA following this protocol was quite pure and ready for sensitive conventional PCR reactions.This is an economic,efficient,and timesaving method.Moreover,crude DNA extracted by other methods can be also further purified with this new way.尽管人们采用多种提取方法乃至使用商品试剂盒,但从森林土壤中获得纯品DNA仍然是比较困难的事情。本文经多次研究探索提出了一种经济有效的森林土壤粗品DNA进一步纯化方法。此方法由两步组成:1.利用提取缓冲液将提取的粗品DNA溶解,经氯仿/异戊醇去除杂质后,再用异内醇沉淀析出DNA;2.使用普通凝胶回收试剂盒回收柱进一步纯化DNA。结果表明:经第一步纯化后,82%-91%腐殖酸杂质被除去。经第二步纯化后,残留的少量腐殖酸杂质被去除干净。经上述连续的两步纯化后获得的DNA非常纯净,可直接用于对抑制物非常敏感的常规PCR反应。本研究报道的DNA进一步纯化方法有效、经济且省时。此外,采用其它各种提取方法获得的土壤粗品DNA均可使用本方法进一步纯化。图4表2参15。

关 键 词：crude DNA forest soils PURIFICATION 

分 类 号：S714[农业科学—林学]