Abstract
We describe here a protocol for obtaining clones containing sequences present in low copy-number from genomic DNA where moderately and highly repeated sequences predominate. Specific chromosomal regions can be targeted by using deletion or addition line material. We have used this protocol to identify a sequence which has been deleted in both the tetraploid and hexaploid wheat mutants for the homoeologous chromosome pairing locus.
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- Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [DOI] [PubMed] [Google Scholar]
- Flavell R. B., Bennett M. D., Smith J. B., Smith D. B. Genome size and the proportion of repeated nucleotide sequence DNA in plants. Biochem Genet. 1974 Oct;12(4):257–269. doi: 10.1007/BF00485947. [DOI] [PubMed] [Google Scholar]
- Kohne D. E., Levison S. A., Byers M. J. Room temperature method for increasing the rate of DNA reassociation by many thousandfold: the phenol emulsion reassociation technique. Biochemistry. 1977 Nov 29;16(24):5329–5341. doi: 10.1021/bi00643a026. [DOI] [PubMed] [Google Scholar]
- Kunkel L. M., Monaco A. P., Middlesworth W., Ochs H. D., Latt S. A. Specific cloning of DNA fragments absent from the DNA of a male patient with an X chromosome deletion. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4778–4782. doi: 10.1073/pnas.82.14.4778. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rimpau J., Smith D., Flavell R. Sequence organisation analysis of the wheat and rye genomes by interspecies DNA/DNA hybridisation. J Mol Biol. 1978 Aug 15;123(3):327–359. doi: 10.1016/0022-2836(78)90083-9. [DOI] [PubMed] [Google Scholar]