Evaluation of a decontamination protocol for hair shafts before mtDNA sequencing.
Mitochondrial DNA (mtDNA) sequencing is a powerful and sensitive method to identify the donor of shed hairs found at a crime scene. Because of the low amounts of DNA in shed hair and the sensitivity of PCR, contaminating cells (e.g. saliva, blood), sometimes present on these hairs, will be co-amplified. This will result in ambiguous sequencing results and might even lead to erroneous exclusions of suspects. We have evaluated a strategy for effectively removing saliva and blood contamination from hair samples. Unambiguous mtDNA results were obtained by incubating the hair samples in a differential lysis buffer (which contains no DTT) prior to DNA extraction. Since the nuclear DNA of the hair root is affected, this procedure should be restricted to hair shaft proportions.
Jehaes
E
E
Center for Human Genetics, University of Leuven, Belgium.
Gilissen
A
A
Cassiman
J J
JJ
Decorte
R
R
D001769:Blood; D016172:DNA Fingerprinting; D017931:DNA Primers / Q000737:chemistry; D004272:DNA, Mitochondrial / Q000032:analysis*; D003666:Decontamination / Q000379:methods*; D005554:Forensic Medicine; D006197:Hair / Q000737:chemistry*; D006801:Humans; D016133:Polymerase Chain Reaction; D012463:Saliva; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004272:DNA, Mitochondrial