Purpose: The purpose of today’s lab was to run our gels from the PCR we made last week from Chelex and V4 primers. We also began discussing and planning our poster design.

Procedure:

1. Obtain gel made last week
2. Fill gel electrophoresis box to create a thin layer of 1x TAE solution over the gel
3. Add 5 μL of DNA ladder
4. Add 10 μL of PCR products
5. Label wells
6. Run gels at 100 volts for 30 minutes
7. Place gels on UV light and take picture
8. Enter data in the Excel sheet
9. Brainstorm for poster design

Results:

Well 1: DNA ladder

Well 2: Group 2 negative PCR

Well 3: Group 2 positive PCR

Well 4: Group 2 eDNA PCR

Well 5: Group 1 negative PCR

Well 6: Group 1 positive PCR

Well 7: Group 1 eDNA PCR

Well 8: none

• Both groups had eDNA in the correct spots.

Conclusion: The Chelex protocol with V4 primers seems to be working about 50% of the time between the two classes. This is good because the number of base pairs present allows metabarcoding to be done which was our original goal at the beginning of the semester.

Storage: We stored our PCR tubes in the same purple container at the front of the room. Our tubes are in row D4, spots 1-3. Spot 1 is negative, spot 2 is positive, and spot 3 is eDNA.