Williams Eletra, Lin Meng-Han, Harbison Sallyann, Fleming Rachel
School of Chemical Sciences, The University of Auckland, Auckland, New Zealand; ESR, Private Bag 92021, Auckland, New Zealand.
ESR, Private Bag 92021, Auckland, New Zealand.
Forensic Sci Int Genet. 2014 Mar;9:85-92. doi: 10.1016/j.fsigen.2013.11.007. Epub 2013 Dec 6.
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.
信使核糖核酸分析正成为法医学中体液鉴定的常用方法，但当存在来自多个个体的细胞混合物时存在缺点。一种能够识别和分离此类细胞混合物的方法将简化下游分析。为此，我们开发了一种新型的RNA悬浮荧光原位杂交（RNA S-FISH）方法，使用针对角蛋白10（KRT10）信使核糖核酸的锁核酸（LNA）探针，该探针适合作为上皮细胞的潜在标志物。由于法医样本的样本量可能受到限制，该方法专注于在保持信号强度的同时尽量减少细胞损失。此外，我们已经证明，从使用激光显微切割分离的150个RNA S-FISH标记细胞中获得完整的DNA图谱是可能的。
