Removal Of Soil Polymerase Chain Reaction Inhibitors By Addition Of Powdered Activated Charcoal | 4735
Journal of Biotechnology & Biomaterials
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Polymerase chain reaction (PCR) represents fast and reliable DNA amplification method applied in many scientific fields.
However, its sensitivity to presence of various PCR inhibitory substances can lead to false negative results. Humic substances
(HS), as major component of soil organic matter, are commonly found in DNA prepared from biological material exposed to
environment. Due to their highly complex chemical composition, soluble HS readily co-purify with DNA and are difficult to
remove without additional, often laborious treatments to obtain DNA suitable for PCR. The objective of the present study was
to investigate removal of soil PCR inhibitors using powdered activated charcoal (PAC). The data show that soluble HS could be
reduced up to 12-folds, based solely on change in temperature and time of adsorption. Complete PCR inhibition occured in 90%
of unpurified samples, while 45% of PAC-purified samples were successfully amplified. In addition, optimized PAC-purification
resulted in minimal loss of DNA and DNA quantity and quality obtained using this simple method and commercially available
ultrafiltration column were compared.
Lucija Barbaric graduated in Biotechnology from Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia. Currently she works
at Forensic Science Centre ?Ivan Vucetic? and pursuing her PhD degree in Molecular Biology from Faculty of Science, University of Zagreb.
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