Lab 7: PCR amplification of DNA
Lab 7: PCR amplification of DNA
The purpose and objective:
The objective of lab is setting up the Polymerase Chain Reaction (PCR) by using our DNA sample to amplify a piece of DNA sample and discussing the scientific poster design.
Date: 02/28/19
Materials: DNA sample, 10% bleach, ice box, D.I. water, tube, primer, 2X Taq Mix, micropipettor, pipette tip
Procedures:
- Wear gloves and clean the desk with 10% bleach;
- Place an empty tube, water, DNA sample, and V4 Primer in a box with ice;
- Add 9µl water first and add 9µl DNA sample to create the 1:10 dilution and mix the solution with micropipettor;
- And then, take two tubes which have already containing 12.5µl 2X Taq Mix;
- Label the control and treatment;
- For the control tube, add 1µl primers (10µM) and 11.5µl water and mix them;
- For the treatment tube, add 1.7µl DNA sample, 1µl primers (10µM), 9.8µl water and mix them;
- Make sure close all tubes and pipette tip boxes when not in use.
Data:
| Component | Volume (Control Tube) | Volume (Soil DNA tube) |
| 2X Master mix | 12.5 | 12.5 |
| DNA Template | 0 | 1.7 |
| Primers (10µM) | 1 | 1 |
| Water | 11.5 | 9.8 |
| Total Volume | 25µl | 25µl |
Storage:
We put the DNA sample, water, primers and tubes back to the box with ice.
Conclusion and the future goal:
In the lab, we learned lots of information about PCR and set up PCR by using our DNA sample successfully. Also, discussed about the outline of Poster. Moreover, we will run Agarose Gel and PCR one more time in the future. And I hope that we could analyze our DNA sample through PCR and learn more about the diversity of ciliates.