A strategy for co-analysis of microRNAs and DNA

MicroRNAs (miRNAs, 18-25 bases in length) are small, non-coding RNAs that regulate gene expression at the post-transcriptional level. The small size makes them more stable than conventional mRNA as a biomarker identifying body fluids in degraded or compromised samples. Compared to mRNA, however, due to the very short length of miRNAs, it is a challenge to design proper primers to achieve miRNAs/DNA co-extraction and co-analysis. Here we describe the design of a specific linear RT primer for the reverse transcription reaction and a pair of PCR primers to be used in the endpoint PCR reaction for each miRNA marker, and presented a strategy for co-analysis of a set of miRNAs and DNA on the same extract using the same detection platform. A set of 4 miRNA markers (miR214 as menstrual blood marker, miR451a as venous blood marker, miR888 and miR891a as semen markers) and a DNA STR multiplex kit were co-analyzed by capillary electrophoresis. The result demonstrated that the strategy of co-analysis of miRNAs/DNA could not only identify the body fluid, but also obtain a STR profile for the same sample.