Chang Lab:Notebook/CBE/08/148/2008/12/13

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LOG BOOK

Bacteria Culture:

  • Take 5ml from the overnight 10ml Ecoli culture and mixed with 150ml of broth.
  • Incubate this for another 5 hour.

Biofilm Optimization Preparation: (Trial 1)

  • After 5 hours, the incubated Ecoli culture is diluted to obtained the McFarland standard (0.257 @ 600nm)
    • 0.22ml of Ecoli Culture : 2.78ml LB broth gives a relative close reading at 0.258
  • The Ecoli culture is then centrifuge for about 10minutes
  • Pippete out the supernatant of LB Broth, the Ecli sendiment is then mixed with the previously prepare M9 medium and is matched with the Mc Farland Standard
    • 0.5ml of Ecoli culture: 2.5ml of M9 medium gices the closes reading at 0.265
  • Using multipipette, 230 uL of adjusted(according to Mcfarland standard) inoculum is 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.

Notes:

  • M9 medium is not homogeneous, we will be trouble-shooting this problem by finding another recipe for preparation of M9 medium.