A technique for nucleic acid in situ hybridization to polytene chromosomes of mosquitoes in the Anopheles gambiae complex

A sensitive, simple, and reproducible in situ hybridization technique for the detection and precise localization of specific nucleic acid sequences on chromosomes of members of the Anopheles gambiae complex is described. Modifications of the in situ hybridization technique are described that allow simultaneous hybridization of several probes with the chromosomes on a single slide and the multiple use of a single chromosome preparation for several different probes hybridized successively on the same slide. Examples are shown that illustrate the utility of the technique for localization of both single copy and repeated sequences in both polytenized euchromatin and centromeric heterochromatin.