A Rapid and Economical Method for Efficient DNA Extraction from Diverse Soils Suitable for Metagenomic Applications

被引：44

作者：

Devi, Selvaraju Gayathri ^{[1
]}

Fathima, Anwar Aliya ^{[1
]}

Radha, Sudhakar ^{[1
]}

Arunraj, Rex ^{[1
]}

Curtis, Wayne R. ^{[2
]}

Ramya, Mohandass ^{[1
]}

机构：

[1] SRM Univ, Dept Genet Engn, Kattankulathur, Tamil Nadu, India

[2] Penn State Univ, Dept Chem Engn, University Pk, PA 16802 USA

来源：

PLOS ONE | 2015年 / 10卷 / 07期

关键词：

SEDIMENT SAMPLES; PURIFICATION PROCEDURES; MOLECULAR DIVERSITY; BACTERIAL DIVERSITY; COASTAL GUJARAT; SALINE HABITATS; NUCLEIC-ACIDS; GLASS POWDER; PCR; ENVIRONMENT;

D O I：

10.1371/journal.pone.0132441

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

A rapid, cost effective method of metagenomic DNA extraction from soil is a useful tool for environmental microbiology. The present work describes an improved method of DNA extraction namely "powdered glass method" from diverse soils. The method involves the use of sterile glass powder for cell lysis followed by addition of 1% powdered activated charcoal (PAC) as purifying agent to remove humic substances. The method yielded substantial DNA (5.87 +/- 0.04 mu g/g of soil) with high purity (A(260/280): 1.76 +/- 0.05) and reduced humic substances (A(340): 0.047 +/- 0.03). The quality of the extracted DNA was compared against five different methods based on 16S rDNA PCR amplification, BamHI digestion and validated using quantitative PCR. The digested DNA was used for a metagenomic library construction with the transformation efficiency of 4 X 10(6) CFU mL(-1). Besides providing rapid, efficient and economical extraction of metgenomic DNA from diverse soils, this method's applicability is also demonstrated for cultivated organisms (Gram positive B. subtilis NRRL-B-201, Gram negative E. coli MTCC40, and a microalgae C. sorokiniana UTEX#1666).

引用

页数：16