Purpose/Objective of the Lab:

• The purpose of today’s lab was to design a protocol for the metabarcoding of soil  ciliates to explore the ciliate biodiversity.

General Work flow:

• Ciliate Isolation 
• DNA Extraction
• PCR/Gel Electrophoresis 
• Metabarcoding
• Analysis of Sequencing Data 

Procedure:

1. We added 8 mL Ludox HS 40 to a 15 mL conical tube (falcon tube was used in this procedure
2. We quickly injected 2 mL of distilled water to the top of the Ludox mixture.
3. The whole class gave their falcon tubes with the components to the instructor and we went upstairs to centrifuge them in a swinging bucket for 15 minuts at 4300 x g(earth’s gravitaional pull)
4. We carefully removed the tube and used a pippette to remove the cell layer under the water layer.
5. We then trandferred 3-10 microliter drops to a concavity slide and counted the cells using a 100x magnification.
6. After we  observed these drops under the microscope, we then prepared the cells for DNA extraction by first labelling our tubes and spining them in the microfuge for about 1 minute.
7. We then stored the microfuge tube with the pellet on the bottom in the freezer.

Results:

• The mass of the falcon tube before centrifufing it was approximately 21.1 grams.
• Unfortunately, we were not able to locate any ciliates under the microscope.

Conclusion:

• Our falcon tube did not look the same compared to other groups because we quickly added the water into the tube making it mix with the ludox and soil sample that was already in the falcon tube. Even though there was not a very distinct line in our tube, we were still able to locate some organic material after it was centrifuged.