Journal article
Snapback SSCP analysis: Engineered conformation changes for the rapid typing of known mutations
Human Mutation, Vol.11(3), pp.252-258
1998
Abstract
Several approaches may be applied to detect known mutations, including restriction enzyme cleavage, allele-specific oligonucleotide (ASO) hybridization or amplification, dideoxy fingerprinting, and direct DNA sequencing. All these approaches require several extra steps after PCR and may involve radioactive isotopes, time-consuming hybridization, template purification, or digestion steps. The ease and simplicity of the SSCP test make it a popular choice for mutation detection, but a significant limitation is that some DNA changes will not alter the overall conformation of either single strand and are thus not amenable to SSCP typing. We describe Snapback SSCP to genotype normal and mdx mice (an animal model of Duchenne muscular dystrophy) that previously could not be differentiated by conventional SSCP analysis. A snapback primer was designed with additional bases at the 5′ terminus, which were complementary to the normal sequence flanking the mdx mutation and used under the original amplification conditions. Each single strand of these snapback PCR products now had one terminus capable of re-annealing or “snapping back” to the normal sequence but not the mdx mutation. In this manner, a conformation change was engineered into the normal strand that could be readily distinguished from the mdx allele on a SSCP gel. This approach could be applied to the routine screening of other known mutations that are not amenable to detection by simple SSCP analysis.
Details
- Title
- Snapback SSCP analysis: Engineered conformation changes for the rapid typing of known mutations
- Authors/Creators
- S.D. Wilton (Author/Creator) - Centre for Neuromuscular and Neurological DisordersK. Honeyman (Author/Creator) - The University of Western AustraliaS. Fletcher (Author/Creator) - Centre for Neuromuscular and Neurological DisordersN.G. Laing (Author/Creator) - UWA Centre for Medical Research
- Publication Details
- Human Mutation, Vol.11(3), pp.252-258
- Publisher
- John Wiley & Sons Inc.
- Identifiers
- 991005540336407891
- Copyright
- © 1998 Wiley-Liss, Inc.
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
UN Sustainable Development Goals (SDGs)
This output has contributed to the advancement of the following goals:
Source: InCites
Metrics
33 Record Views
InCites Highlights
These are selected metrics from InCites Benchmarking & Analytics tool, related to this output
- Collaboration types
- Domestic collaboration
- Citation topics
- 1 Clinical & Life Sciences
- 1.255 Musculoskeletal Disorders
- 1.255.628 Duchenne Muscular Dystrophy
- Web Of Science research areas
- Genetics & Heredity
- ESI research areas
- Molecular Biology & Genetics