IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/09

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Prep for UV Induction
Reagents to be prepared Autoclaved both at 250 degrees F for 25 minutes
 * Soft Agar - 0.65% Agar
 * Making 400mL, probably only need 26mL tomorrow
 * Recipe: 8.5g LB Agar / 400mL water
 * Hard Agar - 1.5% Agar
 * Making 400mL, probably only need 180mL tomorrow
 * Recipe: 18.5g LB Agar / 400mL water


 * Made 1/10 dilution of LB Broth
 * Recipe: 3mL LB Broth + 27mL sdH20

Began overnight cultures of 8325 + RN4220

All overnight cultures were done in 13mL test tubes with 3mL LB Broth. Two overnight controls were used.

Place in incubator @ 37 degrees C, 220 RPM @ 1745.

Tomorrow:
 * 2 independent UV induction protocol run throughs
 * Day 2 of Biofilm protocol
 * Refine Biofilm protocol

Gel verification on ligation mixes from Aug 05 2010
Gel orientation: Results:
 * Changes: 1% agarose gel
 * Samples: 1,2,3,4
 * Machine conditions: 0.5x TBE buffer, 100V, 45min

Transformation

 * 1) Testing competency
 * 2) Transform in pBAD (Amp-resistant)
 * 3) Changes: add 1uL of pBAD to competent cells
 * 4) Transforming ligation mixes (1,3,4 - all Amp-resistant)
 * 5) Changes: add 6uL of each to competent cells

- Both incubate for 1 hour at 37C - Both spread on Amp plates - Plates 1,3,4pBAD in 37C at 1540 Vicki Ma 02:55, 11 August 2010 (EDT)