Abstract
dc:description.abstract<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>
Degree
thesis:*- Name thesis:degree_name
- Master of Science (MS)
- Level thesis:degree_level
- Open Access Thesis
- Discipline thesis:degree_discipline
- Biology
- Year dc:date.available
- 2022
Author and committee
dc:creator, dc:contributor.*- Author dc:creator
-
- Mishler, Jeanmarie
- Contributors dc:contributor
-
- Anne Casper, Ph.D.
- Robert Winning, Ph.D.
- Andrew Ross, Ph.D.
Subjects
dc:subject × 2Identifiers
dc:identifier.*- Repository record dc:identifier
- https://commons.emich.edu/theses/1174
- OAI identifier oai:identifier
- oai:commons.emich.edu:theses-2552