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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.
Asunto(s)

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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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