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...

Ausführliche Beschreibung

Gespeichert in:

Bibliographische Detailangaben
Veröffentlicht in:	Applied and Environmental Microbiology 2010-09, Vol.76 (17), p.5995-6000
Hauptverfasser:	Mettel, Carsten, Kim, Yongkyu, Shrestha, Pravin Malla, Liesack, Werner
Format:	Artikel
Sprache:	eng
Schlagworte:	Biological and medical sciences Chromatography Chromatography, Ion Exchange Exonucleases - metabolism Fluorescence in situ hybridization Fundamental and applied biological sciences. Psychology Hydrogen-Ion Concentration Metagenomics - methods Methods Microbiology Mitochondria Molecular Weight Nucleic Acid Hybridization Ribonucleic acid RNA RNA, Messenger - chemistry RNA, Messenger - genetics RNA, Messenger - isolation & purification Sepharose Soil Microbiology Soils Time Factors
Online-Zugang:	Volltext
Tags:	Tag hinzufügen Keine Tags, Fügen Sie den ersten Tag hinzu!

Beschreibung
Zusammenfassung:	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.
ISSN:	0099-2240 1098-5336 1098-6596
DOI:	10.1128/aem.03047-09