Removal of humic substances from soil DNA using aluminium sulfate

Direct extraction of soil DNA has become essential for the study of soil microorganisms. Humic substances co-extracted during DNA retrieval is a big problem because it greatly inhibits the enzymes involved in manipulating DNA. Popular commercial kits available for soil DNA extraction are unable to o...

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Bibliographic Details
Published in:Journal of microbiological methods 2006-08, Vol.66 (2), p.217-222
Main Authors: Dong, Dexian, Yan, An, Liu, Haiming, Zhang, Xuehong, Xu, Yuquan
Format: Article
Language:English
Subjects:
Alum Compounds - chemistry
Aluminium sulfate
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
DNA extraction
DNA, Bacterial - isolation & purification
Extracellular DNA
Flocculation
Fundamental and applied biological sciences. Psychology
Humic substance
Humic Substances - microbiology
Hydrogen-Ion Concentration
Microbiology
Soil Microbiology
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Summary:Direct extraction of soil DNA has become essential for the study of soil microorganisms. Humic substances co-extracted during DNA retrieval is a big problem because it greatly inhibits the enzymes involved in manipulating DNA. Popular commercial kits available for soil DNA extraction are unable to overcome this problem. Here we report an effective protocol for the removal of humic substance from soil DNA. The protocol involves flocculation of the humic substance by excessive Al 3+, then removal of superfluous Al 3+ via pH adjustment and finally release of soil microbial DNA by SDS lysis. This technique is superior to that employed by the UltraClean Soil DNA Kit and can be applied to a wide variety of soils.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2005.11.010