| 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
|
| Project Status |
Completed (Fiscal Year 2017)
|
| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
|
|---|
| Keywords | mRNA / 抽出・精製方法 / マルチプレックスRT-PCR / RNA |
|---|
| 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.
|
