Procedure

1. Seed each vessel with K. rhaeticus stock and Yamanaka media at 1:10 ratio, respectively. Remove the air filter on the end of the tubing and replace it with the media filter. Using the syringe, pump in the appropriate amount of Yamanaka + Glucose Media. Removing the media filter, use the micropipette to pipette in K. rhaeticus cells. Do not pump in K. rhaeticus cells through filters. Place the air filter back onto the tubing to act as an air vent when media and cells are in the vessels.

Rect 1 : Media + Culture: 400mL of space

Rect 2: Media + Culture: 250mL of space

Cylin 1: Media + Culture: 100mL of space

Cylin 2: Media + Culture: 50mL of space

Above is the given amount of space that that should be filled by media and culture for each container. This is half of the overall volume of each container, so that the media level doesn't touch the tubing that will be used to sterilely put in media and culture, and cause irregular BC growth.

2. Incubate at 30C for 10-14 days.

3. Follow harvesting and drying protocol to collect BC and mass. Record masses in the below table.

Results

4. Compare the BC mass to Total Volume Ratios with each other, to see if the square container design will work just as well as the circular container.

Observations (9/14)

6. 500mL bottle had a clump of BC that appears to be growing, under the white spot of that clump were a lot of wispies which I think might be contamination...

7. 250mL bottle had a little more BC in it, and had a more obvious pellicle start to form, because there appeared to be a whole thin sheet forming over the top

8. large flat pyrex had a lot of BC growin in it. The coloration was appropriate (darkers, phlemmy color) but it was still splotchy

9. Small taller pyrex had less bc clumps