2/22/18
The first task conduct PCR on our DNA sample and our two controls. The Environmental DNA came from an amalgamation of DNA taken from the best samples after last week’s process. The positive control used DNA taken from a culture of Paramecium cells. The negative control had no DNA added to it.
These are the final compositions of our 3 tubes for the PCR reaction:
| Component | Negative Control | Positive Control | Environmental DNA |
| 2X Master Mix | 12.5 μl | 12.5 μl | 12.5 μl |
| DNA template | 0.0 μl | 5.0 μl | 5.0 μl |
| Primers | 1.0 μl | 1.0 μl | 1.0 μl |
| Water | 11.5 μl | 6.5 μl | 6.5 μl |
| Total volume | 25.0 μl | 25.0 μl | 25.0 μl |
While the PCR reaction was taking place we prepared the agarose gel in preparation for gel electrophoresis. The gel was made by mixing 0.6 g of agarose with 40 mL of TAE in an erlenmeyer flask. The flask was microwaved at 7 power for 1 minute and 20 seconds. Next it was placed in a 55 °C bath for 5 minutes. After the bath, 2 μl of ethidium bromide were added to the solution. The solution was then poured into a gel rack and was left at room temperature to set.