Bioassay Optimization in Identification of Transgenic Mice
Location
CSU Ballroom
Start Date
9-4-2012 12:00 AM
End Date
9-4-2012 11:30 AM
Student's Major
Biological Sciences
Student's College
Science, Engineering and Technology
Mentor's Name
Marilyn Hart
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Description
Transgenic mice play a critical role in biology by providing a tool to investigate gene structure, function, and expression. Because of their genetic homology with humans, studying the interactions of proteins in heart cells of transgenic mice can provide insight into potential applications to treat cardiovascular disease. In this study, a PCR assay was refined and optimized to genotype a line of mice carrying a transgene mutation for Actin Capping Protein (CP). DNA was collected by tail snips, digested overnight, and then extracted by standard techniques. Optimization of the PCR reaction was carried out through a series of buffer combinations that varied salt concentration and pH, with β-Actin primers being used. Preliminary results indicated that a buffer containing 2.5mM Mg2+ at pH 9 worked best to amplify the β-Actin control product. Further modifications such as annealing temperature and concentration of template DNA may be investigated to further optimize the assay. The development of a dependable, repeatable essay will be important for maintaining the integrity of the transgenic mouse line.
Bioassay Optimization in Identification of Transgenic Mice
CSU Ballroom
Transgenic mice play a critical role in biology by providing a tool to investigate gene structure, function, and expression. Because of their genetic homology with humans, studying the interactions of proteins in heart cells of transgenic mice can provide insight into potential applications to treat cardiovascular disease. In this study, a PCR assay was refined and optimized to genotype a line of mice carrying a transgene mutation for Actin Capping Protein (CP). DNA was collected by tail snips, digested overnight, and then extracted by standard techniques. Optimization of the PCR reaction was carried out through a series of buffer combinations that varied salt concentration and pH, with β-Actin primers being used. Preliminary results indicated that a buffer containing 2.5mM Mg2+ at pH 9 worked best to amplify the β-Actin control product. Further modifications such as annealing temperature and concentration of template DNA may be investigated to further optimize the assay. The development of a dependable, repeatable essay will be important for maintaining the integrity of the transgenic mouse line.
Recommended Citation
Backes, Nichoas. "Bioassay Optimization in Identification of Transgenic Mice." Undergraduate Research Symposium, Mankato, MN, April 9, 2012.
https://cornerstone.lib.mnsu.edu/urs/2012/poster-session-A/17
