PCR amplification of crude microbial DNA extracted from soil.
A rapid, inexpensive, large-scale DNA extraction method involving minimal purification has been developed that is applicable to various soil types. DNA was extracted from 100 g of soil using direct lysis with glass beads and sodium dodecyl sulphate (SDS) followed by polyethylene glycol precipitation, potassium acetate precipitation, phenol extraction and isopropanol precipitation. The crude extract could be used in PCR directed at high-copy number (bacterial small subunit rRNA) and single-copy (fungal beta-tubulin) genes.
Yeates
C
C
Key Centre for Biodiversity and Bioresources, School of Biological Sciences, Macquarie University, Sydney, Australia. cyeates@rna.bio.mq.edu.au
Gillings
M R
MR
Davison
A D
AD
Altavilla
N
N
Veal
D A
DA
D001483:Base Sequence; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D005800:Genes, Fungal; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012988:Soil Microbiology; D014404:Tubulin / Q000235:genetics
D017931:DNA Primers; D004269:DNA, Bacterial; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D014404:Tubulin; D004247:DNA