Improved Technique of Digoxigenin Labeled RNA in situ Hybridization
Korean Journal of Pathology
;
: 98-110, 2001.
Artículo
en Coreano
| WPRIM
| ID: wpr-173558
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.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Parafina
/
Patología
/
Ribonucleasas
/
Autopsia
/
ARN
/
ARN Mensajero
/
Sondas ARN
/
Reacción en Cadena de la Polimerasa
/
Hibridación in Situ
/
Endopeptidasa K
Idioma:
Coreano
Revista:
Korean Journal of Pathology
Año:
2001
Tipo del documento:
Artículo
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