4/12/18

Purpose: The purpose of today’s lab was to put our PCR products in the agarose gels we made last week and perform gel electrophoresis on them. After we ran our gels, we took them upstairs to observe them under UV light. Lastly, we began brainstorming and designing layouts for our posters.

Procedure:

A. Gel Electrophoresis

1. Obtain the gel made last time we met for lab. (For clarification, last week we created a a 1.8% agarose gel).
2. Pour the 1xTAE solution in the box and just barely over the gel.
3. Obtain the tubes with the PCR products from last week from the front of the classroom. (My group: 21-3 +, 21-3 -, 21-3 e; table group: 21-4 +, 21-4 -, 21-4 e).
4. Pipette 10 uL of each solution into the wells of the gel.
5. Obtain a ladder and pipette 5 uL of it into a well.
6. Cover the box and ensure all wires are where they are supposed to be.
7. Run gel electrophoresis for atleast 30 minutes at 100 volts.
8. When done, send a group member upstairs with Dr. Adair to view the gels under UV light.

B. Poster Designing

1. As a group complete the QTMs and discuss the following for your poster:
   • Title
   • Abstract
   • Introduction
   • Methods
   • Results
   • Acknowledgements/bibliography

Results:

The samples were placed in the gel in the following order (right to left): ladder, empty space, 21-4e, 21-4 +, 21-4 -, 21-3 e, 21-3 +, 21-3-.

My table group and I were very pleased with our results! We both had bands in our + DNA control and our eDNA. Our table group probably had the best results as their bands were bright and roughly 450 BP.

Conclusion: Todays lab was fairly easy and relaxed. Yay for seeing bands in our +DNA and eDNA!! Hopefully we have enough time to proceed with Illumina sequencing! If not, this lab has been so much fun and has really helped me develop a growth mindset, an attribute that will be helpful in further research participation.

Future Steps: If time permits, the next step would be to send our PCR products to Illumina to metabarcode our DNA. After, our next steps would be to make our posters and present them at the symposium.

Storage: The 6  PCR product tubes labeled 21-3 +, 21-3 -, 21-3 e, 21-4 +, 21-4 -, 21-4 e were placed back in the box in the front of the classroom. Our gel was placed back in the carrier box after observing it under UV light.