High resolution detection of DNA–RNA hybrids in situ by indirect immunofluorescence

Abstract

WE describe here a new method for the detection of RNA–DNA hybrids in cytological preparations with which we have revealed the locations of hybrid molecules on polytene chromosomes. The critical reagent is an antiserum raised in rabbits against poly(rA)·poly(dT) complexed with methylated bovine serum albumin, originally described by Stollar1. The specificity and resolving power of the indirect immunofluorescence procedure are demonstrated using in situ hybridisation of 5S rRNA (ribosomal RNA) to polytene chromosomes of Drosophila melanogaster as a model system. The method has significant advantages over the autoradiographic procedures2–5 used so far.

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RUDKIN, G., STOLLAR, B. High resolution detection of DNA–RNA hybrids in situ by indirect immunofluorescence. Nature 265, 472–473 (1977). https://doi.org/10.1038/265472a0

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