Lab 7: PCR Amplification
Megan Tran
3/1/19
Objective:
The purpose of this lab was to set up a PCR reaction using the eDNA and to begin thinking about the format and design of our upcoming scientific posters.
Procedure:
PCR:
- Calculate the amount of DNA and water that is needed to perform PCR and set up your treatment tube with the extracted DNA.
- After making the calculations, add 11.5 μl of extracted DNA (a dilution is not necessary with our sample because of the small amount of DNA present).
- Get a separate tube with 12.5 μl of 2X Taq Mix, and label each tube control and treatment appropriately.
- Add 12.5 ul of the 2x Taq mix and 1 ul of the primer to the control tube. Then add 11.5 ul of water to make the total volume equal 25 ul.
- Add 12.5 ul of the 2x Taq mix, 11.5 ul of our DNA sample and 1ul of the primer to the experimental tube.
Data:
| Tube | 1 (Control) | 2 (Extracted DNA) |
| 2x Taq Mix | 12.5 | 12.5 |
| DNA (μl) | 0 | 11.5 |
| 10 μM primers (μl) | 1 | 1 |
| Water (μl) | 11.5 | 0 |
| Total Volume (μl) | 25 | 25 |
ng/ul = 7.1
Storage:
The tubes with the primers, our DNA samples, and water were re-stored in a cooler, and the tube containing the rest of the diluted DNA sample was disposed of.
Conclusion: During this lab we learned the steps necessary to prepare our DNA extracted samples for PCR next week. Through all of these different steps we are taking with our DNA samples, we will be able to sequence them and hopefully be able to contribute to the knowledge of ciliate diversity.
Future Steps: I hope that the next time we perform PCR and gel electrophoresis again that our group will find significantly more DNA in our samples, and I hope that I do well on this upcoming midterm.