{"id":{"repo_id":"vt","oai_identifier":"oai:vtechworks.lib.vt.edu:10919/32325"},"canonical_url":"https://search.dev.ndltd.org/etd/vt/oai:vtechworks.lib.vt.edu:10919/32325","repository":{"repo_id":"vt","name":"Virginia Tech","base_url":"https://vtechworks.lib.vt.edu/oai/request"},"display":{"title":"Development of a SNP Assay for the Differentiation of Allelic Variations in the mdx Dystrophic Mouse Model","abstract":"The 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.","abstract_html":"The 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.","abstract_has_math":false,"creators":["Misyak, Sarah A."],"institution":"Virginia Tech","degree_name":"Master of Science","degree_level":"masters","degree_discipline":"Human Nutrition, Foods, and Exercise","degree_department":"Human Nutrition, Foods, and Exercise","school":null,"contributors":[],"advisors":[],"committee_chairs":["Grange, Robert W."],"committee_members":["Good, Deborah J.","Eisenberg, Arthur","Walker, Richard A."],"year":2008,"date_issued":"2008-04-21","date_published":"2008-04-21","updated_at":"2026-07-22T22:20:41Z","subjects":["Mini-sequencing","DMD","mdx","SNaPshot","ARMS"],"languages":[],"rights":["In Copyright"],"rights_urls":["http://rightsstatements.org/vocab/InC/1.0/"],"identifier_entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["etd-05072008-210505"],"render_values":[{"text":"etd-05072008-210505","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/10919/32325","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.committeechair","label":"Committee Chair","values":["Grange, Robert W."]},{"key":"dc:contributor.committeemember","label":"Committee Member","values":["Good, Deborah J.","Eisenberg, Arthur","Walker, Richard A."]},{"key":"dc:contributor.department","label":"Department","values":["Human Nutrition, Foods, and Exercise"]},{"key":"dc:creator","label":"Author","values":["Misyak, Sarah A."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2014-03-14T20:35:31Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2014-03-14T20:35:31Z","2010-12-22"]},{"key":"dc:date.issued","label":"Date","values":["2008-04-21"]},{"key":"dc:publisher","label":"Institution","values":["Virginia Tech"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Human Nutrition, Foods, and Exercise"]},{"key":"thesis:degree_level","label":"Degree Level","values":["masters"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["Virginia Polytechnic Institute and State University"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Mini-sequencing","DMD","mdx","SNaPshot","ARMS"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:rights","label":"Dc Rights","values":["In Copyright"]},{"key":"dc:rights.uri","label":"Rights URI","values":["http://rightsstatements.org/vocab/InC/1.0/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["etd-05072008-210505"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/10919/32325"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["The 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."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Master of Science"]},{"key":"dc:title","label":"Title","values":["Development of a SNP Assay for the Differentiation of Allelic Variations in the mdx Dystrophic Mouse Model"]}]}],"canonical_facts":{"dc:contributor.committeechair":["Grange, Robert W."],"dc:contributor.committeemember":["Good, Deborah J.","Eisenberg, Arthur","Walker, Richard A."],"dc:contributor.department":["Human Nutrition, Foods, and Exercise"],"dc:creator":["Misyak, Sarah A."],"dc:date.accessioned":["2014-03-14T20:35:31Z"],"dc:date.available":["2014-03-14T20:35:31Z","2010-12-22"],"dc:date.issued":["2008-04-21"],"dc:description.abstract":["The purpose of this study was to develop a SNaPshot® assay to simultaneously discriminate between the dystrophic and wild type (wt) alleles in mdx mice. 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