title: A Rapid and Economical Method for Efficient DNA Extraction from Diverse Soils Suitable for Metagenomic Applications.
abstract: 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 μg/g of soil) with high purity (A260/280: 1.76 ± 0.05) and reduced humic substances (A340: 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 106 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).
authors.affiliation: Department of Genetic Engineering, SRM University, Kattankulathur, Tamilnadu, India.
authors.forename: Selvaraju Gayathri
authors.initials: SG
authors.lastname: Devi
authors.affiliation: Department of Genetic Engineering, SRM University, Kattankulathur, Tamilnadu, India.
authors.forename: Anwar Aliya
authors.initials: AA
authors.lastname: Fathima
authors.affiliation: Department of Genetic Engineering, SRM University, Kattankulathur, Tamilnadu, India.
authors.forename: Sudhakar
authors.initials: S
authors.lastname: Radha
authors.affiliation: Department of Genetic Engineering, SRM University, Kattankulathur, Tamilnadu, India.
authors.forename: Rex
authors.initials: R
authors.lastname: Arunraj
authors.affiliation: Department of Chemical Engineering, Pennsylvania State University, University Park, Pennsylvania, United States of America.
authors.forename: Wayne R
authors.initials: WR
authors.lastname: Curtis
authors.affiliation: Department of Genetic Engineering, SRM University, Kattankulathur, Tamilnadu, India.
authors.forename: Mohandass
authors.initials: M
authors.lastname: Ramya
mesh_terms: D003362:Cost-Benefit Analysis; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D015698:Genomic Library; D006812:Humic Substances / Q000032:analysis / Q000382:microbiology; D056186:Metagenomics / Q000191:economics / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics / Q000302:isolation & purification; D012987:Soil / Q000737:chemistry*; D012988:Soil Microbiology
chemical_list: D004269:DNA, Bacterial; D006812:Humic Substances; D012336:RNA, Ribosomal, 16S; D012987:Soil