Removal of PCR inhibitors from soil DNA by chemical flocculation.
Extracting high-purity DNA directly from soil has become essential for the study of microorganisms in environmental samples. However, many soils contain compounds that inhibit enzymes involved in manipulating DNA. In this study, chemical flocculation using multivalent cations was investigated as a potential method for eliminating soil-based inhibitors during the extraction process. The addition of AlNH(4)(SO(4))(2) during extraction significantly reduced the co-purification of PCR inhibitors with minimal loss of DNA yield.
Braid
Michael D
MD
Ciphergen Biosystems, Incorporated, 6611 Dumbarton Circle, Fremont, CA 94555, USA.
Daniels
Laura M
LM
Kitts
Christopher L
CL
D000534:Alum Compounds / Q000378:metabolism; D002122:Calcium Chloride / Q000378:metabolism; D002412:Cations / Q000819:agonists; D002712:Chlorides; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005290:Ferric Compounds / Q000378:metabolism; D005427:Flocculation; D015636:Magnesium Chloride / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D012988:Soil Microbiology
D000534:Alum Compounds; D002412:Cations; D002712:Chlorides; D004269:DNA, Bacterial; D005290:Ferric Compounds; D015636:Magnesium Chloride; C059726:aluminum ammonium sulfate; D002122:Calcium Chloride; C024555:ferric chloride