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Identification and Separation of Same Gender Mixtures of Various Cell Types Using Interphase FISH Techniques and Laser Microdissection.

机译:使用间期FIsH技术和激光显微切割鉴定和分离各种细胞类型的相同性别混合物。

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The generation of clean, single source genetic profiles from sexual assault and touch evidence cellular mixtures is an enduring challenge in the field of forensics. Evidence of this nature can contain low copy amounts of DNA from mixtures of cell types of various morphologies. The goal of this research was to improve the methods of DNA mixture resolution in the forensic laboratory by improving laser microdissection (LM) techniques. Laser microdissection (LM) has proven to be an effective method for cell mixture separations in the forensic laboratory. While sperm and epithelial cell sexual assault mixtures can easily be separated based upon morphological differences, mixtures of the same cell type are more difficult to separate. The key objective of these studies was to separate cellular mixtures of similar morphology and same gender by using sequence specific fluorescence in situ hybridization (FISH) probes, which are based on the genetic polymorphisms associated with the Duffy and ABO blood groups. The Duffy and ABO blood grouping systems were chosen as the basis for these assays because of the existence of their prevalent polymorphisms throughout multiple populations. Each system has several alleles that can be targeted through a multicolor probe assay. The genetic sequences associated with the variations of each system are conserved and manageable in length. Thirty to three hundred base pair long FISH probes were designed to detect the single nucleotide polymorphisms observed between individuals of blood type A and blood type B. Tyramide signal amplification (TSA) was utilized to increase the sensitivity of FISH when detecting the short FISH probes.

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