Detection of chromosomal inversions using non-repetitive nucleic acid probes
A method and a kit for the identification of chromosomal inversions are described. Single-stranded sister chromatids are generated, for example by CO-FISH. A plurality of non-repetitive, labeled probes of relatively small size are hybridized to portions of only one of a pair of single-stranded sister chromatids. If no inversion exists, all of the probes will hybridize to a first chromatid. If an inversion has occurred, these marker probes will be detected on the sister chromatid at the same location as the inversion on the first chromatid.
1. A kit for detecting inversions in a selected mitotic chromosome, comprising a plurality of non-repetitive probes 20 to 90 bases long, wherein each of the probes in the plurality of probes is single-stranded, unique and identical to at least a portion of a target DNA sequence of one single-standard chromatid of the chromosome, each of the probes having at least one fluorescent label, thereby permitting detection thereof by fluorescence in situ hybridization, and further comprising 5′-bromo-2′-deoxyuridine, dye and nuclease for performing the CO-FISH process to generate single-stranded chromatids from the selected mitotic chromosome.
2. The kit of claim 1 , further comprising reagents suitable for hybridizing the probes to the single-stranded chromatids.
3. A kit for detecting inversions in a selected mitotic chromosome, comprising a plurality of non-repetitive probes 20 to 90 bases long, wherein each of the probes in the plurality of probes is single-stranded, unique and complementary to at least a portion of a target DNA sequence of one chromatid of the chromosome, each of the probes having at least one fluorescent label, thereby permitting detection thereof by fluorescence in situ hybridization, and further comprising 5′-bromo-2′-deoxyuridine, dye and nuclease for performing the CO-FISH process to generate single-stranded chromatids from the selected mitotic chromosome.
4. The kit of claim 3 , further comprising reagents suitable for hybridizing the probes to the single-stranded chromatids.