Fluorescent in situ identification of human marker chromosomes using flow sorting and Alu element-mediated PCR.
A novel approach to the identification of human chromosomes has been developed. Chromosomal in situ hybridization (or "chromosome painting") has been performed using Alu element-mediated PCR products from small quantities (250-500) of flow-sorted normal and abnormal chromosomes. Chromosome paints for various normal chromosomes, including 5, 6, 7, 14, 18, 19, 21, and 22, were generated and shown to be effective in the identification of the appropriate chromosomes. In addition, certain abnormal chromosomes, including a mental retardation-associated deletion chromosome 11 (q22-q23), the products of the constitutional translocation t(11;22), and the CML-associated t(9;22), were used to generate region-specific paints. In each case, the appropriate regions of the chromosomes were highlighted and this strategy is, therefore, well suited to the identification of previously unidentified marker chromosomes. A further direct consequence of this work is that chromosome paints specific for the common aberrant chromosomes, such as the Philadelphia chromosome, can be generated and made widely available. These may find particular use in the analysis of complex or masked chromosomal translocations.
Suijkerbuijk
R F
RF
Department of Human Genetics, University Hospital Nijmegen, The Netherlands.
Matthopoulos
D
D
Kearney
L
L
Monard
S
S
Dhut
S
S
Cotter
F E
FE
Herbergs
J
J
van Kessel
A G
AG
Young
B D
BD
D001483:Base Sequence; D002460:Cell Line; D002874:Chromosome Mapping; D002877:Chromosomes, Human; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D005819:Genetic Markers; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000473:pathology; D008214:Lymphocytes / Q000648:ultrastructure; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D014407:Tumor Cells, Cultured
D005456:Fluorescent Dyes; D005819:Genetic Markers