Rapid method for coextraction of DNA and RNA from natural environments for analysis of ribosomal DNA- and rRNA-based microbial community composition.
A rapid protocol for the extraction of total nucleic acids from environmental samples is described. The method facilitates concomitant assessment of microbial 16S rRNA diversity by PCR and reverse transcription-PCR amplification from a single extraction. Denaturing gradient gel electrophoresis microbial community analysis differentiated the active component (rRNA derived) from the total bacterial diversity (ribosomal DNA derived) down the horizons of an established grassland soil.
Griffiths
R I
RI
Molecular Microbial Ecology Laboratory, Institute of Virology and Environmental Microbiology, CEH-Oxford, Oxford OX1 3SR, United Kingdom.
Whiteley
A S
AS
O'Donnell
A G
AG
Bailey
M J
MJ
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification*; D004275:DNA, Ribosomal / Q000235:genetics* / Q000302:isolation & purification*; D017753:Ecosystem; D004587:Electrophoresis, Agar Gel; D004783:Environmental Microbiology; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification*; D012336:RNA, Ribosomal, 16S / Q000235:genetics* / Q000302:isolation & purification*; D012988:Soil Microbiology
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S