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中国农学通报 ›› 2012, Vol. 28 ›› Issue (12): 78-82.doi: 10.11924/j.issn.1000-6850.2011-3342

所属专题: 生物技术

• 农学 农业基础科学 • 上一篇    下一篇

石油污染土壤总DNA的提取

曹小妮 张亚平   

  • 收稿日期:2011-11-10 修回日期:2012-01-20 出版日期:2012-04-25 发布日期:2012-04-25
  • 基金资助:

    国家自然科学基金项目

Extraction of Total DNA from Petroleum-contaminated Soil

  • Received:2011-11-10 Revised:2012-01-20 Online:2012-04-25 Published:2012-04-25

摘要:

寻找一种可获得高浓度、大片段的石油污染土壤微生物总DNA的提取方法。分别采用SDS的细胞裂解法、溶菌酶和SDS裂解法、实验室改进法、试剂盒法提取石油污染土壤总DNA。研究结果表明4种方法都可获得石油污染土壤微生物总DNA,但每种方法在提取量和纯度上有一定差异;实验室改进法提取的DNA产率是最高的,试剂盒法提取DNA纯度最高。实验室改进法获得的微生物总DNA,稀释20倍可直接用于PCR扩增。土壤样品经过一定的预处理后提取的DNA片段大小在23100 bp,实验室改进法提取土壤微生物总DNA产率、纯度较好,不经过纯化可直接用于PCR扩增及其他分子生物学实验的操作。

关键词: 产量, 产量

Abstract:

The aim was to seek a method of high concentration, large fragments of soil microbial total DNA from oil polluted soil. The genomic DNA was isolated from petroleum-contaminated soil by SDS-based cell lysis, SDS-enzyme method, improved SDS-enzyme method, kit method, respectively. The results showed that all of the four methods could extract microbial DNA from petroleum-contaminated soil, but there were some differences in extraction and purity; extracted rate of DNA the laboratory improved method was better than other methods, the purity of DNA the kit method was the highest. Twenty times diluted DNA which extracted by the improved method could be used for PCR. After a certain amount of soil samples of DNA extracted about 23 kb size, the extracted total microbial DNA was nice in extraction and purity, the improved method without purification could be used to PCR amplification and other molecular biology operations.

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