The application of microwave denaturation in comparative genomic hybridization.
Comparative genomic hybridization (CGH) is a powerful tool for analyzing unbalanced chromosomal rearrangements in a variety of tissues. However, reproducibility of the technique is poor. We have developed an alternative protocol involving microwave denaturation of the metaphase chromosome preparations prior to the hybridization step. The advantage of this method for CGH is the retention of the morphology of the chromosomes and hence an improved chromosome banding pattern. Furthermore, it results in a consistently strong hybridization which is not dependent on the batch of lymphocytes used to obtain the metaphase chromosome spreads. This procedure has also proved to be applicable to nucleic acid hybridizations in general. The protocol, its application and the results of this method in CGH is discussed. Furthermore preliminary results of this method in paint and DNA probe hybridizations to chromosome spreads and to RNA in tissue sections are presented.
de Meulemeester
M
M
Institute of Human Genetics, University of Amsterdam, Academic Medical Centre, Netherlands.
Vink
A
A
Jakobs
M
M
Hermsen
M
M
Steenman
M
M
Slater
R
R
Dietrich
A
A
Mannens
M
M
D004273:DNA, Neoplasm / Q000032:analysis; D016678:Genome; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008872:Microwaves; D009691:Nucleic Acid Denaturation; D015203:Reproducibility of Results; D009396:Wilms Tumor / Q000235:genetics / Q000473:pathology
D004273:DNA, Neoplasm