Establishment of the protocol for mRNA quantification after FACS.
| Project/Area Number |
26460676
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Osaka University |
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Principal Investigator |
Takano Toru 大阪大学, 医学系研究科, 講師 (00263236)
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| Project Period (FY) |
2014-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | Flow cytometry / 癌幹細胞 / FACS / Flow Cytometry / 幹細胞 / mRNA / SPIA / Flow Cytometry / リニア増幅法 |
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| Outline of Final Research Achievements |
In our previous studies, we established a method to analyze cells collected by fluorescence-activated cell sorting (FACS), named mRNA quantification after FACS (FACS-mQ). In FACS-mQ, cells are labeled with a fluorescence dyes, and then cells sorted by FACS are examined by analyzing their gene expression profile. Using the previous FACS-mQ protocol, it was not possible to apply some methods, such as amplification of RNAs using T7 promoter, due to RNA degradation. In this study, we tried to modify the FACS-mQ protocol to prevent RNA degradation during FACS. Addition of RNase inhibitor and DTT to some buffers used in FACS-mQ resulted in a high RNA integrity number after FACS. Furthermore, no significant differences in the images of flow cytometry were observed between specimens with or without RNase inhibitor and DTT. Thus, this modification to the FACS-mQ protocol assured the quality of RNAs in cells recovered by FACS.
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Report
(4 results)
Research Products
(23 results)
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[Journal Article] Relationship between dose of antithyroid drugs and adverse events in pediatric patients with Graves’ disease2017
Author(s)
Yasuda, K., Miyoshi, Y., Tachibana, M., Namba, N., Miki, K., Nakata, Y., Takano, T.
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Journal Title
Clinical Pediatric Endocrinology
Volume: 26
Issue: 1
Pages: 1-7
DOI
NAID
ISSN
0918-5739, 1347-7358
Related Report
Peer Reviewed
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[Journal Article] Optimization of recovery and analysis of RNA in sorted cells in mRNA quantification after fluorescence activated cell sorting.2016
Author(s)
Maeda, T., Date, A., Watanabe, M., Hidaka, Y., Iwatani, Y., Takano, T.
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Journal Title
Ann Clin Lab Science
Volume: 46
Pages: 571-577
Related Report
Peer Reviewed / Acknowledgement Compliant
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[Journal Article] Expression analysis of stemness genes in a rat thyroid cell line FRTL52015
Author(s)
Shimasue, A., Yamakawa, N., Watanabe, M., Hidaka, Y., Iwatani, Y., Takano, T.
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Journal Title
Exp Clin Endocrinol Diabetes
Volume: 123
Issue: 01
Pages: 48-54
DOI
Related Report
Peer Reviewed
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