2013 Fiscal Year Final Research Report
Detecting mRNA with low expression levels on tissues using in situ RT-PCR Hybridization method: a pilot study
| Project/Area Number |
22590634
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Legal medicine
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| Research Institution | Shimane University |
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Principal Investigator |
TOGA Tomoko 島根大学, 医学部, 技術専門職員 (20403462)
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| Co-Investigator(Kenkyū-buntansha) |
TAKESHITA Haruo 島根大学, 医学部, 教授 (90292599)
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| Project Period (FY) |
2010-04-01 – 2013-03-31
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| Keywords | in-situ RT-PCR / mRNA / in situ Hybridization / 川本法 |
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| Research Abstract |
In order to detect small-quantity mRNA on a tissue section, reverse transcription PCR was given to create cDNA, next PCR was given to target mRNA. But amplification of the target gene by PCR was not completed.
Then assumed to be stored efficiently RNA, using frozen sections fixed on the adhesive film (Kawamoto method) was applied to in situ Hybridization. Although the adhesive film could bear the protocol of in situ Hybridization in intensity, tissue sections often fall in the washing, and the dyeing residual substance remained in the film. In result, clear target's mRNA localization could not be found.
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