IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/14

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Biofilm Growth Protocol Day 1

 * Began 5mL TSB O/N cultures of 8325-4 and RN4220
 * Eric and Melody each made 1 control tube, 1 8325-4 tube and 1 RN4220 tube
 * Tubes went into the incubator at 1510 @ 37°C w/o shaking
 * Should be removed by 0910 tomorrow (Sunday August 15th) morning

Biofilm Growth Protocol Day 4

 * Removed Test 1 and Test 2 from the bio safety cabinet
 * Made a 0.1% solution of crystal violet
 * 25mL sdH20
 * 25μL 100% crystal violet
 * Performed the procedure as stated until air dry step
 * Let stand next to flame to air dry for 2 hours
 * Due to the plate reader being reserved for the next very long time, no readings can be taken until Monday August 16th @ 1630
 * Test 1 and Test 2 plates will be stored in the bio safety cabinet until then

Autoclaved

 * 6 boxes of 1000μL tips
 * 3 boxes of 10μL tips
 * 2 things of microcentrifuge tubes
 * 1 jar of PCR tubes

Glassware

 * Bleached all test tubes that we knew needed to be disposed of
 * Rinsed everything already in bleaching sink, except for some tubes that had been left w/o bleaching

Organized

 * The bench
 * Falcon tubes

Misc
Jason

Staph Competent cells Test (made July 15)- (Rafael's protocol)
 * Mix 1: 70ul C.cell + 20ul (constitutive promoter + GFP plasmid )
 * Mix 2: 70ul C.cell + 15ul (constitutive promoter + GFP plasmid )
 * Transferred 60 ul of each mix into two electroporation cuvettes
 * Electroporation setting (100ohm, 2.3kva, 2.1ms)
 * Plated on chloramphenicol LB plates
 * Added dissolved NaCl (0.1g/ml dH2O) to each plate
 * Dry near flame
 * Incubate ON at 37C for 48 hours (to be taken out on Monday)

Made 2*40ml Chloramphenicol antiobitics
 * 2.72 g / 80ml 100% ethanol
 * Stored in -20 in falcon tubes (to be aliquoted)
 * Protocol used: (http://openwetware.org/wiki/Endy:Preparing_Antibiotic_Stocks)


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