title: The development of a method of suspension RNA-FISH for forensically relevant epithelial cells using LNA probes.
abstract: Messenger RNA profiling is becoming a common method for body fluid identification in forensic science but there are disadvantages when cell mixtures are present from more than one individual. A method that could identify and separate such cell mixtures would simplify downstream analysis. To do this, we have developed a novel method of RNA suspension-fluorescent in situ hybridization (RNA S-FISH) using a locked nucleic acid (LNA) probe for the keratin 10 (KRT10) mRNA that is suitable as a potential marker for epithelial cells. As sample size may be restricted in forensic samples, this method has focused on minimizing cell loss whilst maintaining signal strength. Furthermore, we have shown that it is possible to obtain full DNA profiles from 150 RNA S-FISH labeled cells isolated using laser microdissection.
authors: School of Chemical Sciences, The University of Auckland, Auckland, New Zealand; ESR, Private Bag 92021, Auckland, New Zealand.
authors: Eletra
authors: E
authors: Williams
authors: ESR, Private Bag 92021, Auckland, New Zealand.
authors: Meng-Han
authors: MH
authors: Lin
authors: ESR, Private Bag 92021, Auckland, New Zealand.
authors: Sallyann
authors: S
authors: Harbison
authors: ESR, Private Bag 92021, Auckland, New Zealand. Electronic address: Rachel.fleming@esr.cri.nz.
authors: Rachel
authors: R
authors: Fleming
mesh_terms: D016172:DNA Fingerprinting / Q000379:methods*; D004847:Epithelial Cells / Q000737:chemistry*; D005260:Female; D005819:Genetic Markers; D006801:Humans; D006824:Hybridization, Genetic; D017404:In Situ Hybridization, Fluorescence; D053550:Keratin-10 / Q000235:genetics; D061346:Laser Capture Microdissection; D009061:Mouth Mucosa / Q000166:cytology; D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D012333:RNA, Messenger / Q000302:isolation & purification*; D017423:Sequence Analysis, RNA; D014621:Vagina / Q000166:cytology
chemical_list: D005819:Genetic Markers; C507493:KRT10 protein, human; D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D012333:RNA, Messenger; C477371:locked nucleic acid; D053550:Keratin-10
keywords: Body fluid identification; Epithelial cells; FISH; Forensic; mRNA