A test project to establish a highly sensitive in situ molecular hybridization method using fluorescent latex beads.
| Project/Area Number |
05557017
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Experimental pathology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
SUGIYAMA Taketoshi Kyoto Univ., Faculty of Medicine Professor, 医学部, 教授 (20030851)
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| Co-Investigator(Kenkyū-buntansha) |
HAGA H Kyoto Univ., Faculty of MedicineResearch Associate, 医学部, 助手 (10252462)
OSAKA M Kyoto Univ., Faculty of MedicineResearch Associate, 医学部, 助手 (20252463)
TAKAHASHI R Kyoto Univ., Faculty of MedicineAssociate Professor, 医学部, 助教授 (60144565)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1993: ¥3,900,000 (Direct Cost: ¥3,900,000)
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| Keywords | latex beads / in situ molecular hybridization / FITC / ビーズ / in situ法 / 蛍光 / 分子雑種 |
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| Research Abstract |
A test was planned to establish a highly sensitive in situ molecular hybridization method using fluorescent letex beads. In this plan, we considerd to conjugate avidine-conjugated FITC-labeled latex beads of various size to the biotin-labeled DNA hybridized in situ to the gene to be demonstrated. We could use beads of 50,100, and 500 A of diameter.
Since the interval of each biotin-labeled DNA base, which was inserted each 20 of bases, was 60 A,the beads of 50 A of diameter was only applicable to the present purpose. However, due to the detachment of the conjugated beads during washing process, faint fluorescence compared to FITC itself (10A) and difficulty in quantitative evaluation, FITC-avidin was concluded to be superior to FITC-labeled latex beads conjugated with avidin. However, the present project clarified the mechanism of in situ hybridization at the dye particle lavel.
Two other experiments were added to this project. Assignment of erythropoietin recepter gene of the rat to #8 chromosome and DNA degradation in biological speciment in atmospheric oxygen in the presence of cellular lipids were two other results obtained in the present studies.
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Report
(3 results)
Research Products
(7 results)
