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Merck

Extraction of mRNA from soil.

Applied and environmental microbiology (2010-07-14)
Carsten Mettel, Yongkyu Kim, Pravin Malla Shrestha, Werner Liesack
ABSTRAKT

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.

MATERIAŁY
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Sigma-Aldrich
Q Sepharose Fast Flow, preswollen, 45-165 μm (wet), exclusion limit ~4,000,000 Da
Sigma-Aldrich
Q Sepharose High Performance, preswollen, 24-44 μm (wet), average exclusion limit ~4,000,000 Da