Improved Technique of Digoxigenin Labeled RNA in situ Hybridization
Korean Journal of Pathology
; : 98-110, 2001.
Article
em Ko
| WPRIM
| ID: wpr-173558
Biblioteca responsável:
WPRO
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. CONCLUSION: 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.
Palavras-chave
Texto completo:
1
Base de dados:
WPRIM
Assunto principal:
Parafina
/
Patologia
/
Ribonucleases
/
Autopsia
/
RNA
/
RNA Mensageiro
/
Sondas RNA
/
Reação em Cadeia da Polimerase
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Hibridização In Situ
/
Endopeptidase K
Idioma:
Ko
Revista:
Korean Journal of Pathology
Ano de publicação:
2001
Tipo de documento:
Article