Improved Technique of Digoxigenin Labeled RNA in situ Hybridization
Korean Journal of Pathology
;
: 98-110, 2001.
Article
in Korean
| 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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Paraffin
/
Pathology
/
Ribonucleases
/
Autopsy
/
RNA
/
RNA, Messenger
/
RNA Probes
/
Polymerase Chain Reaction
/
In Situ Hybridization
/
Endopeptidase K
Language:
Korean
Journal:
Korean Journal of Pathology
Year:
2001
Type:
Article
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