title: STR profiling of epithelial cells identified by X/Y-FISH labelling and laser microdissection using standard and elevated PCR conditions.
abstract: During the investigation of allegations of sexual assault, samples are frequently encountered that contain DNA from a female and a male donor. These may represent contributions of DNA from the complainant and potentially, the offender. Many semen stained samples successfully undergo DNA analysis and interpretation using a differential extraction method that separates sperm from the epithelial cells present in the stain. However, for those mixed cell samples that contain only epithelial cells, separation of any male cells from female cells is problematic. This paper describes the application of fluorescent in situ hybridisation (FISH) for the gender identification of epithelial cells and subsequent recovery of target cells using laser microdissection (LMD). The profiling results obtained from samples of known cell numbers using the Identifiler™ multiplex at standard 28-cycle PCR conditions and, when cell numbers are low, the SGM Plus™ multiplex at elevated 34-cycle PCR conditions (also known as Low Copy Number DNA analysis (LCN)) are described.
authors.affiliation: ESR, Private Bag 92-021, Auckland, New Zealand.
authors.forename: Laura
authors.initials: L
authors.lastname: Lynch
authors.affiliation: ESR, Private Bag 92-021, Auckland, New Zealand.
authors.forename: Amelia
authors.initials: A
authors.lastname: Gamblin
authors.affiliation: ESR, Private Bag 92-021, Auckland, New Zealand. Electronic address: sue.vintiner@esr.cri.nz.
authors.forename: Sue
authors.initials: S
authors.lastname: Vintiner
authors.affiliation: ESR, Private Bag 92-021, Auckland, New Zealand.
authors.forename: Joanne L
authors.initials: JL
authors.lastname: Simons
mesh_terms: D004247:DNA / Q000032:analysis / Q000235:genetics; D004847:Epithelial Cells / Q000737:chemistry* / Q000166:cytology; D005260:Female; D053803:Forensic Genetics / Q000379:methods*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D061346:Laser Capture Microdissection / Q000379:methods*; D008297:Male; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction / Q000379:methods; D012732:Sex Determination Analysis / Q000379:methods*; D012742:Sex Offenses / Q000331:legislation & jurisprudence*
chemical_list: D004247:DNA
keywords: Epithelial cells; Fluorescent in situ hybridisation; Forensic DNA; Identifiler; Laser microdissection; Low Copy Number