DNA Isolation and Analysis



DNA Isolation and Analysis

We extract DNA from the cotton swabs using MoBio's PowerSoil™ DNA Isolation kit and stored the eluted DNA samples at -20°C. In hopes to develop a cost-effective and more effective procedure to quantify Bd, we are using traditional PCR to work on detecting the presence of amphibian DNA before any downstream applications. By determining if we isolated amphibian DNA, which should be present since swabbing dislodges epithelial cells, we can rule out one category of false negatives during Bd analysis. That category is Bd false negatives due to lack of DNA in the sample. Amphibian screening samples are run using a MJ Mini™ Gradient Thermal Cycler. We use primers developed by Kocher et al. 1989 that amplify a specific region of ribosomal DNA found in all amphibians (For more on primers, read Kocher et al. 1989). Samples that test positive for amphibian DNA are ready to test for the presence of Bd.

Inside of a MJ Mini™ (Top) and a autoradiogram of one of our gels (Bottom)



Photo credits can be found on acknowlegements page
This site was designed and created by G. Bomar as part of a summer undergraduate research program at Davidson College.

Bd Detection and Data Analysis






















For comments or questions contact G. Bomar: grbomar@davidson.edu
Davidson College,  Davidson,  North Carolina 28035.