Effects of RNA purification procedures on mRNA-based body fluid identification

2017 Fiscal Year Final Research Report

• Project/Area Number: 26460895
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Legal medicine
• Research Institution: National Research Institute of Police Science
• Principal Investigator: Akutsu Tomoko 科学警察研究所, 法科学第一部, 室長 (50356151)
• Co-Investigator(Kenkyū-buntansha): 櫻田 宏一 東京医科歯科大学, 大学院医歯学総合研究科, 教授 (10334228)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Keywords: mRNA / 抽出・精製方法 / マルチプレックスRT-PCR

Outline of Final Research Achievements

To ascertain the effects of purification procedures on the mRNA-based body fluid identification, comparative analyses of the yield and quality of total RNA were performed between different purification procedures. As a result, silica column can purify higher-quality RNA; however, the sensitivity of the real-time RT-PCR analysis was higher in the magnetic beads-purified samples caused by the relative efficiency of extracting short-length RNA from degraded samples. We also show that the quantification of relative levels of body fluid-specific genes could be influenced by the purification procedure. Although use of high-quality RNA is generally required for gene expression analysis, the relevance of short RNA fragments cannot be ruled out in forensic samples. Then, a multiplex RT-PCR assay, which can be applied regardless of the purification procedure, was developed as a robust tool for body fluid identification. Each amplicon could be detected and quantified by chip electrophoresis.

Free Research Field

法生物学