Direct detection of expanded trinucleotide repeats using DNA hybridization techniques
- Univ. of Toronto (Canada); and others
Recently, unstable trinucleotide repeats have been shown to be the etiologic factor in several neuropsychiatric diseases, and they may play a similar role in other disorders. To our knowledge, a method that detects expanded trinucleotide sequences with the opportunity for direct localization and cloning has not been achieved. We have developed a set of hybridization-based methods for direct detection of unstable DNA expansion. Our analysis of myotonic dystrophy patients that possess different degrees of (CTG){sub n} expansion, versus unaffected controls, has demonstrated the identification of the trinucleotide instability site without any prior information regarding genetic map location. High stringency modified Southern blot hybridization with a PCR-generated trinucleotide repeat probe allowed us to detect the DNA fragment containing the expansion in myotonic dystrophy patients. The same probe was used for fluorescent in situ hybridization and several regions of (CTG){sub n}/(CAG){sub n} repeats in the human genome were detected, including the myotonic dystrophy locus on chromosome 19q. These strategies can be applied to directly clone genes involved in disorders caused by unstable DNA.
- OSTI ID:
- 134280
- Report Number(s):
- CONF-941009-; ISSN 0002-9297; TRN: 95:005313-1013
- Journal Information:
- American Journal of Human Genetics, Vol. 55, Issue Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
- Country of Publication:
- United States
- Language:
- English
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