Extraction of DNA from forensic-type sexual assault specimens using simple, rapid sonication procedures.
Sonication procedures for the extraction of DNA from forensic-type semen specimens have been developed, which, when compared to currently utilized sperm DNA extraction techniques, are simple, rapid and result in comparable DNA yields. Sperm DNA extraction by sonication was performed on whole semen, seminal stains, buccal swabs and post-coital specimens. Ultrasound disruption of sperm cells and their ultimate release of cellular DNA has been conducted in the presence of sperm wash buffers followed by organic extraction or Chelex 100 with little or no compromise to DNA quality, quantity or amplifiability. Two advantages of sonication over currently used forensic techniques to extract sperm DNA include 1) sperm DNA extraction that occurs within five minutes of sonication compared with an hour or greater for water bath incubations in classic enzyme digestion DNA extractions and 2) one less preparatory step with the Chelex/sonication protocol and three less steps with the sonication/organic protocol compared with other procedures thus eliminating potential sample-to-sample cross-contamination. Sperm DNA extracted by optimum sonication procedures was used for forensic HLA DQ alpha typing and restriction fragment length polymorphisms analysis without any adverse effects on typing results.
Crouse
C A
CA
Crime Laboratory, Palm Beach County Sheriff's Office, West Palm Beach, FL 33406.
Ban
J D
JD
D'Alessio
J K
JK
D002610:Cheek; D003075:Coitus; D004247:DNA / Q000302:isolation & purification*; D004229:Dithiothreitol; D019286:Endopeptidase K; D005554:Forensic Medicine; D006683:HLA-DQ Antigens / Q000032:analysis; D006801:Humans; D008297:Male; D012150:Polymorphism, Restriction Fragment Length; D012661:Semen / Q000166:cytology; D012697:Serine Endopeptidases / Q000378:metabolism; D012742:Sex Offenses; D013010:Sonication; D013094:Spermatozoa / Q000737:chemistry*
D006683:HLA-DQ Antigens; D004247:DNA; D012697:Serine Endopeptidases; D019286:Endopeptidase K; D004229:Dithiothreitol