Lindsay and Danielle viewed the image of the latest gel and annotated it. We determined that the 35S promoter primers worked, but the Rubisco primers did not cut. Afterwards, we cleaned the sink and equipment around the lab.
With the final PCR done, the next step was to pour a gel and load the samples onto it to see if the PCR worked. Dr. Todd already poured the gel when we came in, so we poured TAE buffer into the gel running box and the gel. We removed the well combs and loaded … Continue reading 5/9 Loading/Running the Final Gel
Today Danielle and Jess followed a new DNA extraction protocol in hopes to better the quality of the DNA we get for the PCR reactions. We followed the protocol below: Put in ¼ flake Vortex and grind w pestle Add 300μL 1% saline solution Vortex Microfuge 2 min Take all supernatant and put in 300μL … Continue reading 5/4 New DNA Extraction Protocol
After looking at the results from the previous gel we ran, we think some of the DNA we are using might be degrading. We redid a chelex extraction with cereals A and B. Then, we ran PCR with the new DNA from those two cereals, a negative control (just the primers), and a positive control … Continue reading 5/22 More PCR
Today, we worked with Dr. Todd on the procedure for making a PAGE gel (Polyacrylamide gel electrophoresis) to run our PCR samples in. This was a new skill for our group members and that was somewhat evident. We hit some road blocks due to our inexperience. We used Accogel, 50x TAE and sterile water to … Continue reading 5/1 Making the PAGE Gel
As seen in the gel below, cereals A and B have degraded DNA; however, cereals C, D, and E had good amplification with Rubisco Primers. Unfortunately no primers or amplification appeared in the rows with test cereals and GMO primers. To fix these issues, we will do a new DNA extraction of cereals A and … Continue reading 4/26 Cereals A and B DNA Extraction and PCR Redo