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Journal Article

Extraction of mRNA from Soil

MPS-Authors
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Mettel,  C.
Department of Biogeochemistry, Alumni, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

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Kim,  Y.
Department of Biogeochemistry, Alumni, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

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Shrestha,  P. M.
Department of Biogeochemistry, Alumni, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

/persons/resource/persons254493

Liesack,  W.
Department-Independent Research Group Methanotrophic Bacteria, and Environmental Genomics/Transcriptomics, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

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Citation

Mettel, C., Kim, Y., Shrestha, P. M., & Liesack, W. (2010). Extraction of mRNA from Soil. Applied and Environmental Microbiology, 76(17), 5995-6000. doi:10.1128/aem.03047-09.


Cite as: https://hdl.handle.net/21.11116/0000-0007-C323-5
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.