33P is preferable to 35S for labeling probes used in in situ hybridization

We compared RNA probes labeled with 35S-UTP and 33P-UTP for use in in situ hybridizations. 33P-UTP was readily incorporated into in vitro transcribed RNA, producing 33P-labeled riboprobes of high specific activity. When the 33P- and 35S-labeled riboprobes were compared in in situ hybridizations usin...

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Bibliographic Details
Published inBioTechniques Vol. 15; no. 3; p. 506
Main Authors McLaughlin, S K, Margolskee, R F
Format Journal Article
LanguageEnglish
Published England 01.09.1993
Subjects
Animals
In Situ Hybridization
Isotope Labeling
Phosphorus Radioisotopes
Rats
Rats, Sprague-Dawley
Retina - chemistry
RNA Probes
RNA, Messenger - analysis
Sulfur Radioisotopes
Taste Buds - chemistry
Transcription, Genetic
Transducin - genetics
Uridine Triphosphate - metabolism
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Summary:We compared RNA probes labeled with 35S-UTP and 33P-UTP for use in in situ hybridizations. 33P-UTP was readily incorporated into in vitro transcribed RNA, producing 33P-labeled riboprobes of high specific activity. When the 33P- and 35S-labeled riboprobes were compared in in situ hybridizations using two different tissues, we found that the 33P-labeled riboprobes were less "sticky" than the 35S-labeled riboprobes, giving significantly less nonspecific background hybridization. Because of the low level of background stickiness, it was possible to use ten times more 33P-labeled riboprobe than 35S-labeled riboprobe without appreciably increasing background hybridization. Our findings indicate that, in most cases, 33P is the isotope of choice when labeling probes for in situ hybridizations.
ISSN:0736-6205
1940-9818