Chang Lab:Notebook/CBE/08/148/2008/12/18

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LOG BOOK

 * Took 5 ml from 10-ml overnight E coli culture into 150 ml broth for 5-6 hr incubation at 37 C (inoculum for biofilm optimization trial III)

Biofilm Optimization Trial III
(Temperature at 42 degree was not used as based on the previous trial, the medium in the well gets evaporated)
 * 3 CBDs were prepared to be incubated at 21, 30 and 37 degree (30rpm for 24 hours).
 * Procedure is the same as the previous 2 trials
 * Adjustment according to McFarland Standard
 * For LB:9X (0.3ml broth culture to 2.7ml broth) dilution to match standard.
 * For M9:LB which match the McFarland Standard is prepared for centrifugation. The supernatant LB was then discarded and M9 medium was added.
 * Using a mivropippete,200 uL of adjusted inoculum pipetted into each well of CBD.
 * Columns 1-6: rich M9 Medium
 * Columns 7-12: minimal LB Medium
 * CBD covered with pegged lid. Incubated at desired temperatures at 30 rpm. Will check on 12/19 to score for biofilm growth.

Serial Plating
To be checked after ~24 hrs for colony counting
 * Plated LB-based 1.0 McFarland E coli inoculum at 10^-10 to 10^-20 dilutions to verify cell number.
 * Plated M9-based 1.0 McFarland E coli inoculum at 10^-6 to 10^-10 dilutions to verify cell number.
 * Plated planktonic cells from CBD wells at 10^-2, 10^-4, 10^-6 dilutions for both LB and M9 wells of 21, 30 and 37 C CBDs.


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