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一种高效、经济的方法,可从不同土壤和沉积物中提取适用于生物技术操作的 DNA。

An efficient and economical method for extraction of DNA amenable to biotechnological manipulations, from diverse soils and sediments.

机构信息

Lignocellulose Biotechnology Laboratory, Department of Microbiology, University of Delhi South Campus, New Delhi, India.

出版信息

J Appl Microbiol. 2014 Apr;116(4):923-33. doi: 10.1111/jam.12420. Epub 2014 Jan 6.

Abstract

AIMS

An attempt was made to optimize a new protocol for isolation of pure metagenomic DNA from soil samples.

METHODS AND RESULTS

Various chemicals (FeCl3 , MgCl2 , CaCl2 and activated charcoal) were tested for their efficacy in isolation of metagenomic DNA from different soil and compost samples. Among these trials, charcoal and MgCl2 when used in combination yielded highly pure DNA free from humic acids and other contaminants. The DNA extracted with the optimized protocol was readily digested, amplified and cloned. Moreover, compared with a well-established commercial DNA isolation kit (UltraClean™ Soil DNA Isolation Kit), our method for DNA isolation was found to be economical. This demonstrated that the method developed can be applied to a wide variety of soil samples and allows handling of multiple samples at a given time.

CONCLUSIONS

The optimized protocol developed has successfully yielded pure metagenomic DNA amenable to biotechnological manipulations.

SIGNIFICANCE AND IMPACT OF THE STUDY

A user-friendly and economical protocol for isolation of DNA from soil and compost samples has been developed.

摘要

目的

尝试优化一种从土壤样品中分离纯宏基因组 DNA 的新方案。

方法和结果

测试了各种化学物质(FeCl3、MgCl2、CaCl2 和活性炭)在分离不同土壤和堆肥样品中的宏基因组 DNA 的效果。在这些试验中,当活性炭和 MgCl2 联合使用时,可获得高度纯净的 DNA,无腐殖酸和其他污染物。用优化的方案提取的 DNA 易于消化、扩增和克隆。此外,与一种成熟的商业 DNA 分离试剂盒(UltraClean™土壤 DNA 分离试剂盒)相比,我们的 DNA 分离方法更经济。这表明所开发的方法可以应用于各种土壤样品,并允许在给定时间处理多个样品。

结论

开发的优化方案成功地获得了适合生物技术操作的纯宏基因组 DNA。

研究的意义和影响

开发了一种方便用户使用且经济的从土壤和堆肥样品中分离 DNA 的方案。

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