Chang Lab:Notebook/CBE/08/146/2008/12/13

{| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Project name
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

Entry title
Time started:915am Bacteria Culture:
 * 5ml taken from the overnight 10ml E.coli culture and added to 150ml of broth.
 * Incubated this for 5 hour.

Time started:215pm Biofilm Optimization Preparation: Serial Plating:
 * After 5 hours, the incubated Ecoli culture was diluted to McFarland standard (Abs:0.257 at 600nm).
 * A reading of 0.258 was obtained for 0.22ml E.coli culture : 2.78ml LB broth.
 * The Ecoli culture was centrifuge for 10minutes (3000g).
 * Pippeted out the supernatant (LB Broth), the E.coli residue was mixed with M9 medium and adjusted to Mc Farland Standard.
 * A reading of 0.265 was obtained for 0.5ml of E.coli culture: 2.5ml of M9 medium
 * 0.230 ml of adjusted(according to Mcfarland standard) inoculum was pipetted into each well of CBD using a multipipette.
 * 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.
 * Plated 10^-8, 10^-10, and 10^-12 dilutions to verify cell number after 5hr incubation. To be checked after ~24 hrs for colony counting.

Notes
 * M9 medium not homogeneous (could be due to saturation or missing of autoclaving step).
 * Need to check with RPM value. 30 rpm from Tre-hardy et al. 150 RPM used was too fast, may have caused the spillage during biofilm growth.


 * }