PMID: 6170491Jan 1, 1981Paper

Rapid and high resolution detection of in situ hybridisation to polytene chromosomes using fluorochrome-labeled RNA

Chromosoma
J G BaumanE Kubli

Abstract

Fluorochrome-labeled RNA allows the rapid detection of in situ hybrids without the need for long exposure times as in the autoradiographical hybridisation methods. Resolution is high because of the high resolving power of fluorescence microscopy. The application of a previously reported method for the hybrido-cytochemical detection of DNA sequences to polytene chromosomes of Drosophilia is described. The specificity and sensitivity of the method are demonstrated by the hybridisation with polytene chromosomes of 1) rhodamine-labeled 5S RNA, to the 5S rRNA sites of D. melanogaster (56F) and D. hydei (23B), 2) rhodamine-labeled RNA complementary to a plasmid containing histone genes, to the 39DE region of D. melanogaster, 3) rhodamine-labeled D. melanogaster tRNA species (Gly-3 and Arg-2), to their respective loci in D. melanogaster, 4) rhodamine-labeled RNA complementary to the insert of plasmid 232.1 containing part of a D. melanogaster heat shock gene from locus 87C, to D. hydei heat shock locus 2-32A. In the latter instance it was possible to demonstrate the labeling of a double band which escaped unambiguous detection by autoradiography in the radioactive cytochemical hybridisation procedure because of the low topological res...Continue Reading

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Citations

Jan 1, 1993·Diagnostic Cytopathology·F H Sarkar, D W Visscher
Oct 14, 2006·Molecular Genetics and Genomics : MGG·M P García Guerreiro, A Fontdevila
Apr 1, 1991·Genetic Analysis, Techniques and Applications·J A McNeilJ B Lawrence
Jun 26, 1998·Journal of Chromatography. a·H H Heng, L C Tsui
Nov 1, 1987·Quarterly Reviews of Biophysics·Z Kam
Jul 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·P R Langer-SaferD C Ward
Sep 1, 1981·The Histochemical Journal·J G Bauman, P van Duijn
Jan 1, 1996·Progress in Histochemistry and Cytochemistry·J C Alers, H van Dekken
Aug 14, 2002·Mayo Clinic Proceedings·Cindy Pham LorentzGordon W Dewald
Dec 15, 2015·Methods : a Companion to Methods in Enzymology·Joseph G Gall
Jan 1, 1983·Archives of Orthopaedic and Trauma Surgery·J Müller-Färber, J Rehn
Aug 15, 1985·Analytical Biochemistry·J P CoghlanG W Tregear

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