DNA preparation from sexual assault cases by selective degradation of contaminating DNA from the victim

• Alex Garvin ^[1] ; Michel Bottinelli ^[2] ; Mauro Gola ^[2] ; Ario Conti ^[3] ; Gianni Soldati ^[2]
  1. [1] Bureco Corporation
  2. [2] Molecular Diagnostic Laboratory, Division of Forensic Genetics
  3. [3] Laboratory of Forensic Chemistry and Toxicology, Institute of Chemistryand Toxicology

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• Localización: Journal of forensic sciences, ISSN-e 1556-4029, ISSN 0022-1198, Vol. 54, Nº. 6, 2009, págs. 1297-1303
• Idioma: inglés
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• Resumen

  • The standard method to purify sperm DNA from vaginal swabs taken from rape victims is to selectively digest the victim’s epithe-lial cells to solubilize the victim’s DNA, and then separate the soluble DNA from the intact sperm by centrifugation. A different approach to remov-ing the soluble victim’s DNA is to selectively degrade it using a nuclease, DNase I. DNase I reduces the amount of soluble DNA by over 1000-fold,while having virtually no effect on the sperm DNA remaining in the sperm head and inaccessible to the enzyme. Nuclease inactivation and spermlysis then yield a soluble, pure male DNA fraction. An aliquot of soluble DNA is removed prior to nuclease addition to provide the victim’s fraction.Vaginal swabs taken at defined time points following consensual sex and taken from rape victims were processed using the nuclease method or thestandard method and the nuclease method gave superior short tandem repeat profiles