In Situ Hybridization Technique As Applied To Cytogenetics

Abstract

In our experimentation with the in situ hybridization technique we were unable to band chromosomes with the standard Giemsa staining procedure following autoradiography. Banding was achieved using a replication banding technique in which bromodeoxyuridine (BrdU) was incorporated during replication. The background particles and debris on the chromosomal preparations, which had been in situ hybridized, was reduced by maintaining constant agitation during post-hybridization washes. Excessive background streaks, resulting from cracks in the radiography emulsion, were nearly eliminated by heating the emulsion to 50° C while dipping the slides.