A sequential co-extraction method for DNA, RNA and protein recovery from soil for future system-based approaches

Eoin Gunnigle; Jean-Baptiste Ramond; Aline Frossard; Mary Seeley; Don Cowan

doi:10.1016/j.mimet.2014.06.004

A sequential co-extraction method for DNA, RNA and protein recovery from soil for future system-based approaches

J Microbiol Methods. 2014 Aug:103:118-23. doi: 10.1016/j.mimet.2014.06.004. Epub 2014 Jun 11.

Authors

Eoin Gunnigle¹, Jean-Baptiste Ramond¹, Aline Frossard¹, Mary Seeley², Don Cowan³

Affiliations

¹ Centre for Microbial Ecology and Genomics (CMEG), Department of Genetics, University of Pretoria, South Africa.
² Gobabeb Research and Training Centre, Walvis Bay, Namibia.
³ Centre for Microbial Ecology and Genomics (CMEG), Department of Genetics, University of Pretoria, South Africa. Electronic address: don.cowan@up.ac.za.

PMID: 24929037
DOI: 10.1016/j.mimet.2014.06.004

Abstract

A co-extraction protocol that sequentially isolates core biopolymer fractions (DNA, RNA, protein) from edaphic microbial communities is presented. In order to confirm compatibility with downstream analyses, bacterial T-RFLP profiles were generated from the DNA- and RNA-derived fractions of an arid-based soil, with metaproteomics undertaken on the corresponding protein fraction.

Keywords: Co-extraction; Metaproteomics; Nucleic acids; Protein; T-RFLP.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chemical Fractionation / methods
DNA / analysis
DNA / isolation & purification*
Metagenomics
Proteins / analysis
Proteins / isolation & purification*
Proteomics
RNA / analysis
RNA / isolation & purification*
Soil / chemistry*
Soil Microbiology

Substances

Proteins
Soil
RNA
DNA