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 B. We will also redo the PCR reactions with cereals C, D, and E with both the Rubisco and GMO primers.
We followed the same procedure as “Preparing the Negative Control” in the protocol to extract DNA from cereals A and B.
The same master mix as all previous reactions was made, but this time we made it for 10 Rx. Then we created these 8 reactions below:
Then we ran the PCR under the following conditions:
- 95 degrees for 2 mins
- 35 cycles of:
- 95 degrees for 1 min
- 58 degrees for 1 min
- 68 degrees for 2 min
- 68 degrees for 10 mins
- Hold at 4 degrees