Dynamic Molecular Combing (DMC) is a method of making visible single molecules of DNA. The method stretches the molecules, attaching them to a glass coverslip; labels them with an intercalating dye or fluorescent antibodies; and images the molecules with fluorescent microscopy. After combing, fluorescent in situ hybridization (FISH) can be used to inspect regions on the DNA molecule. DMC is especially technically challenging for undergraduate research laboratories. To make the technique accessible to an undergraduate laboratory this project addresses three challenges: pH of buffer solution, NaOH exposure time, and halogenated thymidine exposure time in yeast (Saccharomyces cerevisiae). A serial pH experiment is necessary to determine optimal pH in any given laboratory. The optimal NaOH exposure time is 20 minutes. Despite using flow cytometry to determine the amount of time spent in S phase, the expected correlation between halogenated thymidine exposure and labeled tract length was not found, leaving the third challenge for further optimization.
","abstract_html":"<p>Dynamic Molecular Combing (DMC) is a method of making visible single molecules of DNA. The method stretches the molecules, attaching them to a glass coverslip; labels them with an intercalating dye or fluorescent antibodies; and images the molecules with fluorescent microscopy. After combing, fluorescent in situ hybridization (FISH) can be used to inspect regions on the DNA molecule. DMC is especially technically challenging for undergraduate research laboratories. To make the technique accessible to an undergraduate laboratory this project addresses three challenges: pH of buffer solution, NaOH exposure time, and halogenated thymidine exposure time in yeast (Saccharomyces cerevisiae). A serial pH experiment is necessary to determine optimal pH in any given laboratory. The optimal NaOH exposure time is 20 minutes. Despite using flow cytometry to determine the amount of time spent in S phase, the expected correlation between halogenated thymidine exposure and labeled tract length was not found, leaving the third challenge for further optimization.</p>","abstract_has_math":false,"creators":["Mishler, Jeanmarie"],"institution":null,"degree_name":"Master of Science (MS)","degree_level":"Open Access Thesis","degree_discipline":"Biology","degree_department":null,"school":null,"contributors":["Anne Casper, Ph.D.","Robert Winning, Ph.D.","Andrew Ross, Ph.D."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2022,"date_issued":"2022-01-01T08:00:00Z","date_published":"2022-01-01T08:00:00Z","updated_at":"2026-07-24T02:17:47Z","subjects":["combing","Molecular Biology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://commons.emich.edu/theses/1174","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Anne Casper, Ph.D.","Robert Winning, Ph.D.","Andrew Ross, Ph.D."]},{"key":"dc:creator","label":"Author","values":["Mishler, Jeanmarie"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2023-02-01T08:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Open Access Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science (MS)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["combing","Molecular Biology"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://commons.emich.edu/theses/1174"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Dynamic Molecular Combing (DMC) is a method of making visible single molecules of DNA. The method stretches the molecules, attaching them to a glass coverslip; labels them with an intercalating dye or fluorescent antibodies; and images the molecules with fluorescent microscopy. After combing, fluorescent in situ hybridization (FISH) can be used to inspect regions on the DNA molecule. DMC is especially technically challenging for undergraduate research laboratories. To make the technique accessible to an undergraduate laboratory this project addresses three challenges: pH of buffer solution, NaOH exposure time, and halogenated thymidine exposure time in yeast (Saccharomyces cerevisiae). A serial pH experiment is necessary to determine optimal pH in any given laboratory. The optimal NaOH exposure time is 20 minutes. Despite using flow cytometry to determine the amount of time spent in S phase, the expected correlation between halogenated thymidine exposure and labeled tract length was not found, leaving the third challenge for further optimization.
"]},{"key":"dc:title","label":"Title","values":["The promise and perils of molecular combing"]}]}],"canonical_facts":{"dc:contributor":["Anne Casper, Ph.D.","Robert Winning, Ph.D.","Andrew Ross, Ph.D."],"dc:creator":["Mishler, Jeanmarie"],"dc:date.available":["2023-02-01T08:00:00Z"],"dc:description.abstract":["Dynamic Molecular Combing (DMC) is a method of making visible single molecules of DNA. The method stretches the molecules, attaching them to a glass coverslip; labels them with an intercalating dye or fluorescent antibodies; and images the molecules with fluorescent microscopy. After combing, fluorescent in situ hybridization (FISH) can be used to inspect regions on the DNA molecule. DMC is especially technically challenging for undergraduate research laboratories. To make the technique accessible to an undergraduate laboratory this project addresses three challenges: pH of buffer solution, NaOH exposure time, and halogenated thymidine exposure time in yeast (Saccharomyces cerevisiae). A serial pH experiment is necessary to determine optimal pH in any given laboratory. The optimal NaOH exposure time is 20 minutes. Despite using flow cytometry to determine the amount of time spent in S phase, the expected correlation between halogenated thymidine exposure and labeled tract length was not found, leaving the third challenge for further optimization.
"],"dc:identifier":["https://commons.emich.edu/theses/1174"],"dc:subject":["combing","Molecular Biology"],"dc:title":["The promise and perils of molecular combing"],"thesis:degree_discipline":["Biology"],"thesis:degree_level":["Open Access Thesis"],"thesis:degree_name":["Master of Science (MS)"]},"updated_at":"2026-07-24T02:17:47Z"}