Increasing DNA extraction yield from saliva stains with a modified Chelex method.
Recovery, preservation and analysis of body fluid stains is an important aspect of forensic science. PCR-based typing of DNA extracted from recovered stains is often a crucial method to identify a perpetrator or exclude an innocent suspect. This paper reports an improved method of extracting genomic DNA from saliva stains deposited on human skin in simulated bite mark situations. Results of organic (phenol-chloroform) extraction and Chelex extraction were compared to a modified Chelex method developed by the authors. Modifications include pre-extraction preparation with proteinase K and incubations at 56 degrees C and 100 degrees C plus microconcentration of the solution. Quantification results using the classical Chelex extraction method showed that 31.9 +/- 4.22% of the deposited DNA was recovered, but using the modified Chelex extraction method DNA recovery was increased to 47.7 +/- 6.90%. The quantity and quality of extracted DNA was shown to be adequate for PCR-based typing at two STR loci.
Sweet
D
D
University of British Columbia, Faculty of Dentistry, Vancouver, Canada. dsweet@unixg.ubc.ca
Lorente
M
M
Valenzuela
A
A
Lorente
J A
JA
Alvarez
J C
JC
D001733:Bites and Stings; D004247:DNA / Q000302:isolation & purification*; D005554:Forensic Medicine / Q000379:methods*; D006801:Humans; D012015:Reference Standards; D012463:Saliva
D004247:DNA