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Strategy for Co-Analysis of MicroRNAs and DNA

NCJ Number
248138
Journal
Forensic Science International Genetics Volume: 12 Dated: September 2014 Pages: 24-29
Author(s)
Yan Li; Ji Zhang; Wei Wei; Zheng Wang; Mechthild Prinz; Yiping Hou
Date Published
September 2014
Length
6 pages
Annotation
This study designed a specific linear real-time (RT) primer for the reverse transcription reaction and a pair of PCR primers for the end-point PCR reaction for each microRNA (miRNA) marker and developed a strategy of co-extraction and co-analysis of a set of miRNAs and DNA with STR profiling from the same sample, using the same detection platform.
Abstract
Recently, the analysis of cell-specific mRNA expression has been used for identification of body fluid. MicroRNAs belong to a class of small non-protein coding RNA molecules of 18-25 nucleotides in length that regulate gene expression by targeting mRNA. Since the amount of biological stains encountered in forensic casework is usually limited, the potential advantage of the mRNA-based assay includes the ability to co-extract mRNA/DNA from one sample using the same technical platform, which could prevent sample consumption. MicroRNAs (miRNAs) belong to a class of small non-protein coding RNA molecules of 18-25 nucleotides in length that regulate gene expression by targeting mRNA cleavage and/or translational repression. Like mRNA, miRNAs show tissue-specific expression patterns for different organs, cell types, and/or conditions; however, compared to mRNA, the short length of miRNAs make it a challenge to design proper primers to achieve miRNA/DNA co-extraction and co-analysis. The current project established a miRNA multiplex strategy that uses linear RT primers that can be used to construct large, more accurate miRNA-based body fluid identification assays. After RNA/DNA co-extraction, each sample can now also be co-analyzed for miRNAs and STR markers, using the same fluorescent detection platform. Obtaining body fluid type and DNA STR results from the same sample extract and in parallel saves time and conserves forensic evidence. 2 figures and 30 references