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rdf:type |
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lifeskim:mentions |
umls-concept:C0028953,
umls-concept:C0030415,
umls-concept:C0162327,
umls-concept:C0162788,
umls-concept:C0182400,
umls-concept:C0205155,
umls-concept:C0722666,
umls-concept:C1522472,
umls-concept:C1551341,
umls-concept:C1552858,
umls-concept:C1552923,
umls-concept:C1552924,
umls-concept:C1705191
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pubmed:issue |
4
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pubmed:dateCreated |
1989-10-6
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pubmed:abstractText |
An in situ hybridization technique has been developed for assessing poly(A)+ RNA preservation in routine pathology specimens. The method detects poly-adenylated RNA sequences in tissue sections using a biotinylated polydeoxythymidine (poly d(T)) probe. The probe was prepared from single-stranded 25-30 base oligo d(T) and was biotinylated using the enzyme terminal deoxynucleotide transferase with biotin-11-dUTP and dTTP in the ratio 1:4. The hybridization protocol uses varying concentrations of proteinase K to unmask mRNA sequences and the biotin-labelled hybrids are demonstrated after hybridization under standard conditions by the application of streptavidin and biotinylated alkaline phosphatase. Alkaline phosphatase was visualized using a Fast Red naphthol-capture method and the sections were counterstained with haematoxylin. The results have confirmed that the method is specific for poly(A)+ RNA and shows that poly(A)+ RNA can be demonstrated in routine formalin-fixed sections using non-radioactive techniques with retention of morphology. It also provides a means of optimizing the hybridization conditions for specific mRNA probes and produces a staining pattern demonstrating the relative level of poly(A)+ RNA per cell which may reveal new information about cell activity and tissue function.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Biotin,
http://linkedlifedata.com/resource/pubmed/chemical/Deoxyuracil Nucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/Fixatives,
http://linkedlifedata.com/resource/pubmed/chemical/Formaldehyde,
http://linkedlifedata.com/resource/pubmed/chemical/Oligonucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/Paraffin,
http://linkedlifedata.com/resource/pubmed/chemical/Poly A,
http://linkedlifedata.com/resource/pubmed/chemical/Poly T,
http://linkedlifedata.com/resource/pubmed/chemical/Polydeoxyribonucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/RNA,
http://linkedlifedata.com/resource/pubmed/chemical/RNA, Messenger,
http://linkedlifedata.com/resource/pubmed/chemical/RNA Probes,
http://linkedlifedata.com/resource/pubmed/chemical/biotin-11-dUTP
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pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
0022-3417
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
158
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
279-86
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pubmed:dateRevised |
2008-11-21
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pubmed:meshHeading |
pubmed-meshheading:2475601-Base Sequence,
pubmed-meshheading:2475601-Biotin,
pubmed-meshheading:2475601-Colon,
pubmed-meshheading:2475601-Deoxyuracil Nucleotides,
pubmed-meshheading:2475601-Fixatives,
pubmed-meshheading:2475601-Formaldehyde,
pubmed-meshheading:2475601-Humans,
pubmed-meshheading:2475601-Nucleic Acid Hybridization,
pubmed-meshheading:2475601-Oligonucleotides,
pubmed-meshheading:2475601-Palatine Tonsil,
pubmed-meshheading:2475601-Paraffin,
pubmed-meshheading:2475601-Poly A,
pubmed-meshheading:2475601-Poly T,
pubmed-meshheading:2475601-Polydeoxyribonucleotides,
pubmed-meshheading:2475601-RNA,
pubmed-meshheading:2475601-RNA, Messenger,
pubmed-meshheading:2475601-RNA Probes,
pubmed-meshheading:2475601-Tissue Preservation
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pubmed:year |
1989
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pubmed:articleTitle |
In situ hybridization demonstration of poly-adenylated RNA sequences in formalin-fixed paraffin sections using a biotinylated oligonucleotide poly d(T) probe.
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pubmed:affiliation |
Department of Pathology, Leicester Royal Infirmary, U.K.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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