Novel method for the production of multiple colour chromosome paints for use in karyotyping by fluorescence in situ hybridisation.
The development of 24-colour fluorescence in situ hybridisation (FISH) has led to significant advances in cytogenetic research and offers the potential for automated karyotypic analysis. However, these techniques are not in routine research or clinical use because of limitations in methods of probe preparation. This article presents new probe construction protocols and strategies for multiple-colour karyotyping by chromosome painting, which makes the technique more efficient and may lead to more widespread implementation. We used paints generated by our protocols to demonstrate the presence of a cryptic translocation t(13;11;22) in the paediatric sarcoma cell line RMS 1598.
Roberts
I
I
Department of Pathology, University of Cambridge, United Kingdom.
Wienberg
J
J
Nacheva
E
E
Grace
C
C
Griffin
D
D
Coleman
N
N
D020223:Chromosome Painting / Q000379:methods*; D004273:DNA, Neoplasm / Q000032:analysis; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D004076:Digoxigenin / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007621:Karyotyping / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D012509:Sarcoma / Q000737:chemistry; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D003856:Deoxyuracil Nucleotides; C070202:digoxigenin-11-deoxyuridine triphosphate; D004076:Digoxigenin