Extraction of mRNA from soil.
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.
Mettel
Carsten
C
Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Str. 10, D-35043 Marburg, Germany.
Kim
Yongkyu
Y
Shrestha
Pravin Malla
PM
Liesack
Werner
W
D002852:Chromatography, Ion Exchange; D005092:Exonucleases / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D056186:Metagenomics / Q000379:methods*; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D012685:Sepharose; D012988:Soil Microbiology; D013997:Time Factors
D012333:RNA, Messenger; D012685:Sepharose; D005092:Exonucleases