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

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Biofilm Track

Eric F. + Melody

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

Other

Eric F. + Melody

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