SIMULTANEOUS DETECTION OF CENTROMERE-SPECIFIC PROBES AND CHROMOSOME PAINTING LIBRARIES BY A COMBINATION OF PRIMED IN-SITU LABELING AND CHROMOSOME PAINTING (PRINS-PAINTING)

In situ techniques for the detection of specific chromosomes using cen tromeric probes and the decoration of entire chromosomes using chromos ome painting are well established. However, in the deciphering of comp lex chromosomal aberrations it is valuable to be able to detect the ce ntromere and the entire DNA of a specific chromosome in different colo urs simultaneously on the same metaphase. In this report we describe a combination of the primed in situ labelling (PRINS) technique and chr omosome painting for simultaneous visualization of centromere-specific oligonucleotides and chromosome painting libraries. A key feature is that the denaturation step in the PRINS reaction is sufficient to keep the chromosomes denatured for chromosome painting. This means that PR INS and consecutive chromosome painting can be performed as a single p rocedure (PRINS-painting)