Cross-species chromosome painting |
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Authors: | Rens Willem Fu Beiyuan O'Brien Patricia C M Ferguson-Smith Malcolm |
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Institution: | Centre for Veterinary Science, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 OES, UK. wr203@mole.bio.cam.ac.uk |
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Abstract: | Comparative genomics is an important and expanding field of research, and the genome-wide comparison of the chromosome constitution of different species makes a major contribution to this field. Cross-species chromosome painting is a powerful technique for establishing chromosome homology maps, defining the sites of chromosome fusions and fissions, investigating chromosome rearrangements during evolution and constructing ancestral karyotypes. Here the protocol for cross-species chromosome painting is presented. It includes sections on cell culture and metaphase preparation, labeling of chromosome-specific DNA, fluorescent in situ hybridization (chromosome painting) and image analysis. Cell culture and metaphase preparation can take between 1 and 2 wk depending on the cell culture. Labeling of chromosome-specific DNA is completed in 1 d. Fluorescent in situ hybridization can be completed in a maximum of 4 d. |
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