Abstract
RNA analysis can be used to differentiate body tissues. Different cell types produce different mRNAs depending upon environment, stress, and other activity levels. This method can be used to differentiate semen, blood, saliva, and vaginal secretions present in a sample. This information can be used to determine ratios of body fluids present in a sample. In this experiment, mitochondrial RNA (mtRNA) will be extracted and analyzed from simulated crime scene samples containing body fluids. The RNA will be extracted using either a standard guanidine isothiocyanate-phenol-chloroform extraction or Micro Fast-Track mRNA Isolation kit. Due to the limited sample size, DNA and mRNA are co-extracted from forensic samples. After gently lysing the cells and inactivating RNases, the RNA can be separated from the DNA using affinity chromatography using oligo(dT) cellulose columns to bind the polyA+ RNA under high salt conditions. The rRNA is removed using a low salt buffer and the mRNA is eluted using a very low ionic strngth buffer. The cDNA is synthesized from the extracted mRNA using tissue-specific primers for tissue and fluids differentiation. The identified tissues will be reported.
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