Today, the control primers (the Rubisco plant primers) we designed arrived! We used the five test food DNA and the positive MM for 12 reactions (180μL sterilized H2O, 30μL 10x buffer, 6μL DNTPs, and 1.5μL taq). We are going to test each of the five text foods with Dr. Todd’s GMO primers and our Rubisco primers. Before we could use the Rubisco primers, we had to calculate the dilution of primers for the primary stock and working stock.
We created the 10 PCR reactions below:
We ran the PCR under these conditions:
- 94 degrees for 2 mins
- 40 cycles of:
- 94 degrees for 1 min
- 59 degrees for 1 min
- 72 degrees for 2 min
- 72 degrees for 10 mins
- Hold at 15 degrees
After PCR was completed, the 10 tubes were held at 4 degrees Celsius in a freezer.