Reference gene study for forensic body fluid identification

Abstract

Reference genes are used in forensic body fluid identification studies to normalise data generated during gene expression experiments. The use of reference genes improves the reliability of qRT-PCR. In this study, 10 most common reference genes UCE, TEF, GAPDH, 18S rRNA, ACTB, B2M, B-Actin, OAZ1, RPS 29 and S15 widely used in forensic body fluid identification studies were selected from relevant literature and qPCR efficiency and sensitivity of all the reference genes was tested using SYBR Green detection. Stability was also assayed using samples stored at room temperature for 6 months using Taqman assay probes. All the markers except TEF displayed high sensitivity and were detected down to 25 pg of RNA input. Stability study demonstrates that B2M, ACTB, RPS29, and UCE are ideal markers for normalization in forensic body fluid identification studies. The study confirms that reference genes should be selected only upon adequate validation of their suitability