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- Assessing PCR Inhibition from Humic Substances
Assessing PCR Inhibition from Humic Substances
Auteur :
Matheson
Carney D.,
Gurneyage
Carli,
Esau
Neal,
...[et al.]
Date de publication : 01/03/2010
Type : Article
Thème : Chimie de l’environnement
Couverture :
Maroc
URL Article :
http://www.benthamscience.com/open/toeij/articles/V003/38TOEIJ.pdfURL Source :
http://www.benthamscience.com 
Télécharger le document :
(301,50 kB) assessing_PCR_inhibition_humic_substances.pdfRésumé/Sommaire :
Inhibition remains the greatest methodological challenge in molecular analysis of buried biological remains. Inhibitory compounds associated with soil environments comprise primarily of humic acids and fulvic acids, collectively referred to as humic substances. We examined the sensitivity of 13 DNA polymerases to both humic acids and fulvic acids and the concentration at which successful amplification can be achieved. This research identified that all 13 DNA polymerases tested exhibited inhibition with varying concentrations of humic acids and that 5 out of the 13 DNA polymerase tested exhibited inhibition with varying concentrations of fulvic acid. The most tolerant DNA polymerase to inhibition due to the presence of humic and fulvic acids is DNA polymerase followed by KlenTaq LA DNA polymerase and RealTaq DNA polymerase that were both only inhibited by 11g and 110g of humic acids. In addition, we present the use of size exclusion chromatography to remove small molecular weight humic substance, dramatically increasing the success of molecular analysis on material associated with burial. This research has implications to the fields of environmental microbiology, soil science, forensic science and archaeological science.
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