Microchip-based cell lysis and DNA extraction from sperm cells for application to forensic analysis.
The current backlog of casework is among the most significant challenges facing crime laboratories at this time. While the development of next-generation microchip-based technology for expedited forensic casework analysis offers one solution to this problem, this will require the adaptation of manual, large-volume, benchtop chemistry to small volume microfluidic devices. Analysis of evidentiary materials from rape kits where semen or sperm cells are commonly found represents a unique set of challenges for on-chip cell lysis and DNA extraction that must be addressed for successful application. The work presented here details the development of a microdevice capable of DNA extraction directly from sperm cells for application to the analysis of sexual assault evidence. A variety of chemical lysing agents are assessed for inclusion in the extraction protocol and a method for DNA purification from sperm cells is described. Suitability of the extracted DNA for short tandem repeat (STR) analysis is assessed and genetic profiles shown. Finally, on-chip cell lysis methods are evaluated, with results from fluorescence visualization of cell rupture and DNA extraction from an integrated cell lysis and purification with subsequent STR amplification presented. A method for on-chip cell lysis and DNA purification is described, with considerations toward inclusion in an integrated microdevice capable of both differential cell sorting and DNA extraction. The results of this work demonstrate the feasibility of incorporating microchip-based cell lysis and DNA extraction into forensic casework analysis.
Department of Chemistry, McCormick Road, University of Virginia, Charlottesville, VA 22904, USA.
Joan M
JM
Bienvenue
Natalie
N
Duncalf
Daniel
D
Marchiarullo
Jerome P
JP
Ferrance
James P
JP
Landers
D002021:Buffers; D004247:DNA / Q000302:isolation & purification*; D004867:Equipment Design; D005554:Forensic Medicine / Q000379:methods*; D006801:Humans; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D016133:Polymerase Chain Reaction; D011902:Rape / Q000175:diagnosis; D013094:Spermatozoa / Q000166:cytology*
D002021:Buffers; D004247:DNA