Powdered Activated Carbon: An Alternative Approach to Genomic DNA Purification.
Forensic evidence samples are routinely found as stains on various substrates, which may contain substances known to inhibit polymerase chain reaction (PCR). The goal of this study was to evaluate post-Chelex(®) 100 purification using powdered activated carbon (PAC). Mock crime scene DNA extracts were analyzed using quantitative PCR and short tandem repeat (STR) profiling to test the DNA recovery and inhibitor removal using PAC with those of the Amicon(®) Ultra 100K. For extracted bloodstains on soil and wood substrates, PAC and Amicon(®) Ultra 100K generated similar DNA yield and quality. Moreover, the two methods significantly decreased the concentration of humic substances and tannins compared to nonpurified extracts (p < 0.001). In instances where extracts contained indigo dye (bloodstains on denim), Amicon(®) Ultra 100K performed better than PAC due to improved amplifiability. Efficient adsorption of humic substances and tannins, which are common inhibitors, indicates PAC's potential application in the purification of high-template DNA extracts.
Ivan Vučetić Forensic Science Center, General Police Directorate, Ministry of the Interior of the Republic of Croatia, Ilica 335, 10000, Zagreb, Croatia.
Lucija
L
Barbarić
Ivan Vučetić Forensic Science Center, General Police Directorate, Ministry of the Interior of the Republic of Croatia, Ilica 335, 10000, Zagreb, Croatia.
Ivana
I
Bačić
Tissue Typing Center, University Hospital Center Zagreb, Kišpatićeva 12, 10000, Zagreb, Croatia.
Zorana
Z
Grubić
D001774:Blood Chemical Analysis; D001801:Blood Stains; D002606:Charcoal / Q000737:chemistry; D004247:DNA / Q000302:isolation & purification*; D016172:DNA Fingerprinting; D006801:Humans; D006812:Humic Substances; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D011208:Powders; D013634:Tannins / Q000737:chemistry
D006812:Humic Substances; D011208:Powders; D013634:Tannins; D002606:Charcoal; D004247:DNA
Amicon®Ultra 100K; forensic science; humic substances; polymerase chain reaction inhibition; powdered activated carbon; tannins