title: A strategy for co-analysis of microRNAs and DNA.
abstract: 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.
authors.affiliation: Institute of Forensic Medicine, West China School of Basic Science and Forensic Medicine, Sichuan University, Chengdu 610041, Sichuan, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Institute of Forensic Medicine, West China School of Basic Science and Forensic Medicine, Sichuan University, Chengdu 610041, Sichuan, China.
authors.forename: Ji
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Institute of Forensic Medicine, West China School of Basic Science and Forensic Medicine, Sichuan University, Chengdu 610041, Sichuan, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.affiliation: Institute of Forensic Medicine, West China School of Basic Science and Forensic Medicine, Sichuan University, Chengdu 610041, Sichuan, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Sciences, John Jay College of Criminal Justice, New York, NY 10019, USA.
authors.forename: Mechthild
authors.initials: M
authors.lastname: Prinz
authors.affiliation: Institute of Forensic Medicine, West China School of Basic Science and Forensic Medicine, Sichuan University, Chengdu 610041, Sichuan, China. Electronic address: forensic@scu.edu.cn.
authors.forename: Yiping
authors.initials: Y
authors.lastname: Hou
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D017931:DNA Primers; D019075:Electrophoresis, Capillary; D035683:MicroRNAs / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D035683:MicroRNAs; D004247:DNA
keywords: Capillary electrophoresis (CE); Co-analysis; Multiplex reverse transcription endpoint PCR; microRNAs