NCJ Number
213738
Journal
Journal of Forensic Sciences Volume: 51 Issue: 2 Dated: March 2006 Pages: 266-273
Date Published
March 2006
Length
8 pages
Annotation
This paper describes the development of a microdevice capable of DNA extraction directly from sperm cells for application in analyzing sexual assault evidence.
Abstract
The network of disulfide bonds that compose the sperm nuclear membrane makes commonly used cell lysis buffers ineffective; this requires the addition of a disulfide bond- reducing agent for sufficient cell lysis. This paper first describes modifications to the DNA extraction device that allow for easer, more robust fabrication. This is followed by a description of the development of a dual-function buffer capable of the complete lysis of sperm cells that is also amenable to the effective loading of the lysed cellular material for the extraction of DNA. The homebuilt microchip-solid phase extraction (SPE) apparatus consisted of a Harvard Apparatus model 22 syringe pump with a 250 ml Hamilton gas-tight syringe. The syringe was connected to the inlet reservoir of the microdevice. The microdevice-based extraction approach is described. It produced purified DNA from a crude sample in less than 10 ml volume and in a form suitable for STR analysis. Finally, a method for a microchip-based extraction is described, and the dual lysis/loading buffer is assessed for its effectiveness in achieving rapid, decreased volume, on-chip, sperm cell lysis for potential integration with cell-sorting technology. 2 tables, 5 figures, and 25 references