| Project/Area Number |
04670180
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Human pathology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
SUGIYAMA Taketoshi (1993) Kyoto University Faculty of Medicine, Professor, 医学部, 教授 (20030851)
濱崎 周次 (1992) 京都大学, 医学部, 助手 (50208576)
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| Co-Investigator(Kenkyū-buntansha) |
OSAKA M. Kyoto University Faculty of Medicine, Associate, 医学部, 助手 (20252463)
HAGA H. Kyoto University Faculty of Medicine, Associate, 医学部, 助手 (10252462)
杉山 武敏 京都大学, 医学部, 教授 (20030851)
嶋田 俊秀 京都大学, 医学部, 助手 (00235623)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1993: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | DNA / formalin / restriction enzyme / PCR / lambda-phage / fixation / preservation / 制限・酵素 |
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| Research Abstract |
The effect of formalin fixation on DNA was studied by analysing the effects on lambda-DNA and DNA in animal and autopsied liver tissues.
(1)By over-night fixation, formalin split DNA into smaller pieces especially in acidic and saltless conditions at room temperature, although bufferred formalin (BF) in neutral at 4ﾟC prevented DNA fragmentation. On the other hand, lambda-phage DNA became resistent to various restriction enzymes. This effect was dependent on temperature but not on salt concentration of the formalin solution. Thus, these 2 phenomena was considered to be an independent effect of formalin probably caused by scission of phosphate-ester binding for the former, and by cross-linking between the amino groups of the DNA bases for the latter. Regardless of these effects, PCR analysis was successful with lambda-DNA fixed for 2 weeks in 10% formalin and 4 weeks in BF, and with DNA in liver tissues fixed for 6 months in 10% formalin at room temperature. Thus, fixation in BF at 4ﾟC for a short time was recommended as the best fixation method for preservation of DNA in pathology specimens.
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