Digoxigenin을 부착시킨 RNA in situ Hybridization의 개선된 연구 방법
Improved Technique of Digoxigenin Labeled RNA in situ Hybridization
이석근, 김연숙, 송인선, 이상신, 이영준, 김우호, 지제근,
소속 상세정보
이석근 ( Lee Suk-Keun )
강릉대학교 치과대학 구강병리학교실
김연숙 ( Kim Yeon-Sook )
강릉대학교 치과대학 구강병리학교실
송인선 ( Song In-Sun )
강릉대학교 치과대학 구강병리학교실
이상신 ( Lee Sang-Shin )
강릉대학교 치과대학 구강병리학교실
이영준 ( Lee Young-Joon )
강릉대학교 치과대학 구강병리학교실
김우호 ( Kim Woo-Ho )
서울대학교 의과대학 병리학교실
지제근 ( Chi Je-Geun )
서울대학교 의과대학 병리학교실
KMID : 0357920010350020098
Abstract
Background: A practical RNA in situ hybridization method using digoxigenin labeled RNA probes is described in order to evaluate the technical difficulties and problems in RNA in situ hybridization.
Methods: The paraffin sections, routinely processed in the Pathology Laboratory, were tested for the possibility of RNA in situ hybridization instead of the RNase free paraffin sections, fixed in 4% paraformaldehyde and prepared using RNase protection procedures.
Results: Most of the paraffin sections, fixed in 10% neutral formalin solution in fresh condition, showed relatively good reaction of RNA in situ hybridization, although the necrotic tissue and autopsy specimens showed poor reaction of RNA in situ hybridization. A refixation procedure using a 4% paraformaldehyde solution was evaluated for optimal expression of mRNA in the paraffin sections.
Conclusions: The treatment of 4% paraformaldehyde before the treatment of proteinase K showed better in situ hybridization than did the treatment of 4% paraformaldehyde after the treatment of proteinase K. Also a new Polymerase Chain Reaction (PCR)-based method of RNA probe production showed consistently good results.
키워드
RNA;in situ hybridization;Digoxigenin;Polymerase Chain Reaction
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