| Literature DB >> 20622132 |
Carsten Mettel1, Yongkyu Kim, Pravin Malla Shrestha, Werner Liesack.
Abstract
Here, we report an efficient method for extracting high-quality mRNA from soil. Key steps in the isolation of total RNA were low-pH extraction (pH 5.0) and Q-Sepharose chromatography. The removal efficiency of humic acids was 94 to 98% for all soils tested. To enrich mRNA, subtractive hybridization of rRNA was most efficient. Subtractive hybridization may be followed by exonuclease treatment if the focus is on the analysis of unprocessed mRNA. The total extraction method can be completed within 8 h, resulting in enriched mRNA ranging from 200 bp to 4 kb in size.Entities:
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Year: 2010 PMID: 20622132 PMCID: PMC2935057 DOI: 10.1128/AEM.03047-09
Source DB: PubMed Journal: Appl Environ Microbiol ISSN: 0099-2240 Impact factor: 4.792