RNA-seq of biological fluids for the evaluation of mRNA degradation in relation to sample age
Abstract
Research on the forensic applications of RNA analysis has increased greatly in the last decade. Defined uses of RNA in forensic analysis include the use of RNA to identify tissue type, determine sample age, and play a role in molecular autopsies. Although recent research has indicated many possible forensic applications of RNA analysis, many questions remain concerning the behavior of RNA in degraded and limited samples. Specifically, there remains to be a thorough understanding of the differing patterns and rates of RNA degradation in post-mortem and deposited samples. Thus, choosing suitable RNA markers for evaluating the approximate age of a forensic sample can be problematic. Development of a reliable and accurate molecular assay for the determination of sample age (time-since deposition of a biological sample and/or post-mortem interval) will play a critical role in helping investigators establish the timeline of events that surround a crime. The purpose of this research is to evaluate mRNA degradation in forensically relevant biological sample types (blood, saliva, semen, and vaginal fluid) in order to establish tissue-specific transcriptome (total mRNA) degradation profiles and patterns that may correlate with the age of a sample. Transcriptome sequencing of mRNA isolated from fresh and aged samples (0 days to 360 days old) was performed to evaluate the patterns of mRNA degradation in relation to sample age. Sequencing data was used to determine the pattern and rate of degradation for each individual mRNA transcript in each sample type. Sequencing data indicates that the mRNA population and transcript degradation rates appear to be tissue-specific. The mRNA degradation profiles obtained from this study can be used to determine the transcripts in each sample type that have degradation patterns and rates correlated with sample age.
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- OSU Dissertations [11222]