Chang Lab:Notebook/CBE/08/149/2008/12/13

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 * style="background-color: #FFF"|   Antimicrobial efficacy testing of antibiotic-containing biodegradable nanopolymers against biofilm and planktonic cells
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 * style="background-color: #F2F2F2" align="center"|  |Main Notebook page


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Logs

 * Start 9:15 AM
 * 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 )
 * Prepared LB Broth (400 mL)

Biofilm Optimization (Trial I)

 * Following 1.0 McFarland Std (A600 = 0.257), 5-hr incubated E coli broth adjusted to match A600 value. A 13.6X dilution was required to match standard. [[Image:LB_Broth_Culture_Absorbance.jpg|thumb|right|E. coli in LB dilutions]]
 * For minimal medium M9, E coli broth centrifuged at 3000g for 10 minutes, supernatant LB Broth discarded through careful pipetting, without disturbing sedimented E coli cells. Previously prepared M9 medium used to resuspend E coli cells. Absorbance at 600nm adjusted until it matched 1.0 McFarland Std. Approximately 6X dilution to match standard. [[Image:M9_Culture_Absorbance.jpg|thumb|right|E. coli in M9 dilutions]]
 * Further dilution was again carried out (i.e. 1 inoculum : 29 medium)
 * Using multipipette (8-pronged), 230 uL of adjusted inoculum pipetted into each well of CBD.
 * Columns 1-6: rich LB Medium
 * Columns 7-12: minimal M9 Medium
 * CBD covered with pegged lid. Incubated at 4:00 PM at 37C, 150 rpm. Will check on Monday to score for biofilm growth.

Serial Plating

 * Plated 10^-8, 10^-10, and 10^-12 dilutions to verify cell number after 5-hr incubation. To be checked after ~24 hrs for colony counting.