Taylor Hutcheson

Date:
April 7, 2017

Goals:

• Extract the DNA of dense ciliate culture so as to prepare for the process of gel electrophoresis.

Procedure:

1. Transfer 300 uL – 500 uL of ciliate culture to microcentrifuge.
2. Centrifuge for 5 minutes at 6000 g, discarding supernatant.
3. Incubate for 30 minutes in a 56 C water bath, as this will break open the cells and denature some proteins.
4. Boil for 8 minutes in 100 C water bath.
5. Vortex for 1 minute.
6. Centrifuge for 3 minutes at 16000 g to pellet cellular debris.
7. Transfer supernatant with DNA in solution to microcentrifuge tube, without collecting the pellet.
8. Label this tube with the soil identified (mine was TDH06S17) and place it in an ice box.

Observations:
After checking to see if my ciliates had cultured, I was pleasantly surprised to find that, under 40x magnification with a compound microscope, there were too many ciliates to count, all moving around quite rapidly.

Future Experiments:
After obtaining the DNA solution, the next step will be to prepare PCR reactions and gel electrophoresis so that we can identify the type if ciliate in my soil sample.