BjornsMethods/ExperimentLog

December 14, 2005

 * The first step of my adaptation protocol requires me to reproducibly grow bacteria to a concentration of 5e7 - 2e8 cells/mL  in 1 hour


 * Today I am taking one of my frozen aliquots of bacterial cells (BL21), thawing it, diluting it down to 2 different concentrations, and growing it back up to hopefully find out all the details of what initial concentration of cells will give me the desired concentration at the end of 1 hour.


 * Going to try the following initial volumes of my frozen aliquot: 200uL, 500uL


 * 1) X uL of my Klett 80 (4e8 cells/mL) E. coli in ~9.9 mL LB solution in 125 mL flask
 * 2) Grow in orbital water bath at 37C, 200 rpm for 1 hr (or until back to Klett 80)
 * 3) Put flask on ice
 * 4) Serial dilute to get a 1e6 dilution of the Klett 80 solution and create 4 plates: 1x200uL, 2x100uL, 1x50uL
 * 5) Grow overnight in warm room (37C)

Results:
 * After 1 hr, the 200uL sample was only at Klett 3 and the 500uL sample was at Klett 16. I let the Klett 16 sample continue to grow for another hour at which it was at Klett 36.  At the end of one hour I still proceeded to plate and grow the 500uL sample to practice my technique.

December 15, 2005

 * 1) Remove my plates from the warm room
 * 2) Count colonies and calculate the true concentration of my Klett 16 solution

NOTE: I realized when I came in this morning that I diluted down my Klett 16 solutions as though they were of Klett 80, thus I had very few colonies on my plates this morning. Live and learn ehh???

Today I'm going to repeat yesterday's protocol but with an initial bacteria aliquot volume of 1 mL and 1.5 mL since I was not able to get enough bacteria grown at yesterday's initial concentrations.

RESULTS: After 1 hr the 1 mL sample grew to Klett 18 and the 1.5 mL sample grew to Klett 35. I diluted and plated them to grow overnight in the 37C room so I can count them in the morning.

December 16, 2005
Counted the colonies I grew yesterday and confirmed the following bacterial concentrations:


 * 1.5 mL starting aliquot had 1.08e8 cells/mL
 * 1.0 mL starting aliquot had 7.5e7 cells/mL

These results now let us know how much of the initial cell aliquot to add to the LB solution so we have btw 5e7 - 2e8 cells/mL  after 1 hour of growth.

December 17-19, 2005
Been working on my multi-agent systems simulator all weekend. Its pretty cool ;)