Journal of Biotechnology & Biomaterials
Open Access

Abstract

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.

Biography

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.