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dc.contributor.authorMisyak, Sarahen_US
dc.date.accessioned2014-03-14T20:35:31Z
dc.date.available2014-03-14T20:35:31Z
dc.date.issued2008-04-21en_US
dc.identifier.otheretd-05072008-210505en_US
dc.identifier.urihttp://hdl.handle.net/10919/32325
dc.description.abstractThe purpose of this study was to develop a SNaPshot® assay to simultaneously discriminate between the dystrophic and wild type (wt) alleles in mdx mice. The mdx mouse is an animal model for Duchenne muscular dystrophy (DMD), a severe and fatal muscle wasting disease. To evaluate possible treatments and to carry out genetic studies, it is essential to distinguish between mice that carry the mutant dystrophic or wt allele(s). The current Amplification-Resistant Mutation System (ARMS) assay used to genotype mdx mice is labor intensive and sometimes fails to yield typing results, which reduce its efficiency as a screening tool. An alternative assay based on single nucleotide polymorphism (SNP) extension technology (i.e., SNaPshot®) would be advantageous because its specificity and capability to be automated would reduce the labor involved and increase the fidelity of each assay. A SNaPshot® assay has been developed that provides a robust and potentially automatable assay that discriminates between the wt and dystrophic alleles. The assay has been optimized to use: an undiluted DNA in the PCR, a 0.1 µM PCR primer concentration, a full PCR product for the SNP extension reaction, a 50ºC annealing temperature for the SNP extension in accordance with standard SNaPshot® conditions, and a 0.4 µM concentration of the SNP extension primer. The advantages of the resultant SNaPshot® assay over the ARMS assay include higher fidelity, robustness, and more consistent performance within and among laboratories, and reduced risk of human error.en_US
dc.publisherVirginia Techen_US
dc.relation.haspartSM_thesis7_etdcorrections.pdfen_US
dc.rightsI hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to Virginia Tech or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report.en_US
dc.subjectMini-sequencingen_US
dc.subjectDMDen_US
dc.subjectmdxen_US
dc.subjectSNaPshoten_US
dc.subjectARMSen_US
dc.titleDevelopment of a SNP Assay for the Differentiation of Allelic Variations in the mdx Dystrophic Mouse Modelen_US
dc.typeThesisen_US
dc.contributor.departmentHuman Nutrition, Foods, and Exerciseen_US
dc.description.degreeMaster of Scienceen_US
thesis.degree.nameMaster of Scienceen_US
thesis.degree.levelmastersen_US
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen_US
thesis.degree.disciplineHuman Nutrition, Foods, and Exerciseen_US
dc.contributor.committeechairGrange, Robert W.en_US
dc.contributor.committeememberGood, Deborah J.en_US
dc.contributor.committeememberEisenberg, Arthuren_US
dc.contributor.committeememberWalker, Richard A.en_US
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-05072008-210505/en_US
dc.date.sdate2008-05-07en_US
dc.date.rdate2010-12-22
dc.date.adate2008-06-06en_US


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