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Detecting mRNA with low expression levels on tissues using in situ RT-PCR Hybridization method: a pilot study

Research Project

Project/Area Number 22590634
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Legal medicine
Research InstitutionShimane University

Principal Investigator

TOGA Tomoko  島根大学, 医学部, 技術専門職員 (20403462)

Co-Investigator(Kenkyū-buntansha) TAKESHITA Haruo  島根大学, 医学部, 教授 (90292599)
Project Period (FY) 2010-04-01 – 2013-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2011: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2010: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywordsin-situ RT-PCR / mRNA / in situ Hybridization / 川本法 / in situ RT-PCR
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.

Report

(4 results)
  • 2013 Final Research Report ( PDF )
  • 2012 Annual Research Report
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • Research Products

    (17 results)

All 2013 2012 2011 2010

All Journal Article (6 results) (of which Peer Reviewed: 6 results) Presentation (11 results)

  • [Journal Article] Distribution and haplotype analysis of all the non-synonymous and autoimmunity-related single nucleotide polymorphisms in the human deoxyribonuclease II gene using worldwide populations2013

    • Author(s)
      Kaori Kimura
    • Journal Title

      Legal Medicine

      Volume: 15 Issue: 3 Pages: 157-160

    • DOI

      10.1016/j.legalmed.2012.10.002

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Determination of ABO genotypes by real-time PCR using allele-specific primers2012

    • Author(s)
      Muro T, Fujihara J, Imamura S, Nakayama H, Kimura-Kataoka K, Toga T, Iida R, Yasuda T, Takeshita H
    • Journal Title

      Legal Medcine

      Volume: 14(1) Pages: 47-50

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Genetic and expression analysis of SNPs in the human deoxyribonuclease II: SNPs in the promoter region of the gene reduce its in vivo activity through decreased promoter activity2012

    • Author(s)
      Kaori Kimura
    • Journal Title

      Electrophoresis

      Volume: 33 Issue: 18 Pages: 2852-2858

    • DOI

      10.1002/elps.201200260

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Determination of ABO genotypes by real-time PCR using allele-specific primers2012

    • Author(s)
      Muro T, Fujihara J, Imamura S, Nakamura H, Kimura-Kataoka K, Toga T, Iida R, Yasuda T, Takeshita H
    • Journal Title

      Leg Med (Tokyo)

      Volume: 14(1) Pages: 47-50

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Expression of the prolactin receptor in Japanese breast cancer2011

    • Author(s)
      Nio Y, Iguchi C, Tsuboi K, Itakura M, Nishi T, Hashimoto K, Takamura M, Takeda H, Omori H, Koike M, Sato Y, Endo S, Ogo K, Yamaguchi K, Tsuji M, Toga T, Takeshita H
    • Journal Title

      Shimane Journal of Medidal Science

      Volume: 28(1) Pages: 1-6

    • NAID

      120005586218

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Simultaneous determination of seven informative Y chromosome SNPs to differentiate East Asian, European, and African populations.2011

    • Author(s)
      Muro T, Iida R, Fujihara J, Yasuda T, Watanabe Y, Imamura S, Nakamura H, Kimura-Kataoka K, Yuasa I, Toga T, Takeshita H.
    • Journal Title

      Leg Med

      Volume: 13 Pages: 134-141

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] 熱湯およびズダンブラックB処理法を応用した多重蛍光免疫染色の一例2013

    • Author(s)
      栂とも子, 林芳弘, 山根史嗣
    • Organizer
      第24回生物学技術研究会
    • Place of Presentation
      岡崎コンファレンスセンター(岡崎市)
    • Related Report
      2012 Annual Research Report
  • [Presentation] Development of functional Zinc Oxide nanoparticles for biomedical applications2013

    • Author(s)
      Y.Fujita, H.Hashimoto, M. Tongu, T.Yamada, K.Nishimura, K. Yoshikiyo, H. Akiyoshi, T. Yamamoto, A. Matsumoto, M. Fujii, J. Fujihara, T. Toga, H. Takeshita, T. Isobe
    • Organizer
      The China-Japan Symposium on Nanomedicine
    • Place of Presentation
      Nanjin, China
    • Related Report
      2012 Annual Research Report
  • [Presentation] Investigation of Functional Zinc Oxide Nanoparticles for Biomedical Applications2013

    • Author(s)
      Y. Fujita, H. Hashimoto, M. Tongu, T. Yamada, K. Nishimura, K. Yoshikiyo, H. Akiyoshi, T.Yamamoto, A. Matsumoto, M. Fujii, J. Fujihara, T. Toga, H. Takeshita, T. Isobe
    • Organizer
      7th International Symposiumu on Nanomedicine
    • Place of Presentation
      九州工業大学(北九州市)
    • Related Report
      2012 Annual Research Report
  • [Presentation] ヒト石灰化病変の川本法を応用した非脱灰凍結切片の組織染色2012

    • Author(s)
      栂とも子
    • Organizer
      第23回生物学技術研究会
    • Place of Presentation
      岡崎コンファレンスセンター(岡崎市)
    • Year and Date
      2012-02-17
    • Related Report
      2013 Final Research Report 2011 Annual Research Report
  • [Presentation] The Basic Properties of Fluorescence ZnO Nanoparticles for Medical Usage2012

    • Author(s)
      M.Fujii,T.Toga,H.Hashimoto,H.Akiyoshi,H.Takeshita,Y.Fujita
    • Organizer
      6th International Symposiumu on Nanomedicine
    • Place of Presentation
      くにびきメッセ(松江市)
    • Related Report
      2012 Annual Research Report
  • [Presentation] ヒト石灰化大動脈弁の川本法を応用した非脱灰凍結切片の組織染色2011

    • Author(s)
      栂とも子, 木村かおり, 藤原純子, 高橋節典, 高山公子, 大塚洋輔, 安田年博, 竹下治男
    • Organizer
      第28回日本法医学会学術中四国地方集会
    • Place of Presentation
      川崎医科大学(倉敷)
    • Year and Date
      2011-10-15
    • Related Report
      2011 Annual Research Report
  • [Presentation] しじみ貝の殻を含む全体組織染色-非脱灰凍結標本作成技術(川本法)を応用して2011

    • Author(s)
      栂とも子他
    • Organizer
      第22回生物学技術研究会
    • Place of Presentation
      岡崎コンファレンスセンター(岡崎市)
    • Related Report
      2013 Final Research Report
  • [Presentation] しじみ貝の殻を含む全体組織染色-非脱灰硬組織凍結標本作製技術(川本法)を応用して2011

    • Author(s)
      栂とも子, 木村かおり, 藤原純子, 高橋節典, 高山公子, 阿草哲郎, 大塚洋輔, 竹下治男
    • Organizer
      第22回生物学技術研究会
    • Place of Presentation
      岡崎コンファレンスセンター(岡崎市)
    • Related Report
      2010 Annual Research Report
  • [Presentation] 硬組織における遺伝子発現解析研究への非脱灰凍結切片作成技術(川本法)の応用2010

    • Author(s)
      栂とも子他
    • Organizer
      第57回日本法医学会学術近畿地方集会
    • Place of Presentation
      関西医科大学附属病院(枚方市)
    • Year and Date
      2010-11-13
    • Related Report
      2013 Final Research Report
  • [Presentation] 硬組織における遺伝子発現解析研究への非脱灰凍結切片標本作製技術(川本法)の応用2010

    • Author(s)
      栂とも子, 丸山理留敬, 宇田川潤, 高橋節典, 高山公子, 木村かおり, 藤原純子, 大塚洋輔, 竹下治男
    • Organizer
      第57回日本法医学会近畿地方会
    • Place of Presentation
      関西医科大学附属病院(枚方市)
    • Year and Date
      2010-11-13
    • Related Report
      2010 Annual Research Report
  • [Presentation] 大動脈瘤におけるテネイシンXの高発現と高糖鎖修飾2010

    • Author(s)
      松本健一, 織田禎二, 佐藤和美, 塚本万里枝, 荒内ひとみ, 栂とも子, 進藤正信, 戸塚靖則
    • Organizer
      第42回日本結合組織学会学術大会 第57回マトリックス研究会大会 合同学術集会
    • Place of Presentation
      秋田拠点センター・アルヴェ(秋田市)
    • Related Report
      2010 Annual Research Report

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Published: 2010-08-23   Modified: 2019-07-29  

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