Discipline: Biological Sciences
Subcategory: Ecology
Session: 2
Room: Exhibit Hall
Alana Sudkamp - Pima Community College
Co-Author(s): Gabriel Gonzales, Pima Community College; Brittnee Lacasse, Pima Community College; Sophia Dolberg, Pima Community College
The Tucson Bee Collaborative (TBC) works to further research in the biodiversity of bee populations in and around the Sonoran Desert. The Sonoran Desert is thought to be home to the most biodiverse region for bees in the world but little is known about what species are actually present. My role with the Tucson Bee Collaborative was to identify a bee to the species level through DNA barcoding analysis. My classmates and I generated new DNA barcodes which will help researchers and scientists around the world to better identify bees and their biodiversity. As we identify more samples, we become closer to understanding just how many different species exist in the Sonoran Desert. In doing this, we gained experience with scientific research. The Tucson Bee Collaborative is a partnership between Pima Community College, The University of Arizona, and the Arizona Sonoran Desert Museum.To extract DNA, we removed a tissue sample from our bee specimen and mixed it with Proteinase K to release the DNA molecules, then washed the solution with buffers to purify our DNA sample. After mixing our crude DNA sample with a master mix and centrifuging, we ran our sample in a thermocycler where it underwent denaturing, annealing, and extending. This process amplified our original crude DNA and gave us thousands of copies of that DNA sequence to work with. We used gel electrophoresis as a tool to confirm the success of our PCR step in amplifying our DNA before sending it away for analysis. Our gel electrophoresis revealed a very obvious band perfectly in line with our positive control, right around the 600-700 base pair length. This confirmed that our PCR was successful in amplifying the CO1 gene from our extracted DNA sample. Once our amplified sample was sequenced at a remote facility, we received the chromatograms of our DNA sequences in both the forward and reverse directions. From these sequences we were able to build a consensus sequence, then search in the Barcode of Life Database (BOLD) to determine an identification for our specimen.Our search of our consensus sequence on BOLD revealed the top 19 hits as 100% matches for Diadasia diminuta. This leaves us confident in concluding that the identification of Bee #19 is Diadasia diminuta. This record has been published to BOLD for other scientists to make use of our data. This research was supported by the National Science Foundation under Grant No. 1928400.
Funder Acknowledgement(s): This research was supported by the National Science Foundation under Grant No. 1928400.
Faculty Advisor: Jennifer Katcher, jkatcher@pima.edu
Role: DNA extraction, PCR, gel electrophoresis, bioinformatics to identify bee to species level.