IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/01

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BioFilm track

Obtained 2 strains of ON cultures of Staph RN4220 (From 2 different TSA plates (JG and MW))

  • Prepared 2 sets, 4 beakers of TSB + (10%, 20% w/v glucose );
  • Dilute 1 strain (JG) of staph ON culture (1:100) in 1 set of beakers with TSB + (10, 20 w/v glucose)
  • Diluted 2nd strain (MW) of staph ON culture 1:50 in 2nd set of TSB beakers


Per strain;

  • Tested in Triplicates; 3 wells per glucose concentration
  • number of tests per glucose concentration: 4 (4 sets of triplicates)
  • Total number of tests per glucose concentration: 3*4=12
  • Volume of ON culture + TSB per well = 200ul
  • Total volume of TSB per glucose concentration: 200ul* 12 = 2.4 ml


Total volume of TSB = 2 sets of strains * 2.4ml + 8(200ul, negative controls) =4.8ml + 1.6ml =6.4 ml of TSB

Dilution Factors and Glucose Added

  • 3ml of TSB per glass beaker
  • 7 beakers (1 per glucose concentration per strain + 1 for negative control)
  • 1:100; 30ul ON culture: 3ml TSB
  • 1:50; 60ul ON culture: 3ml TSB
  • Glucose added: (0.3g, 0.6g)

Legend

96 well Plate Growth
Well ColumnStrainStrain:TSBGlucose % (w/v)
AJG1:10010
BJG1:10020
DMW1:5020
EMW1:5010
FControl-10
GControl-10

Additional Notes

  • Vortexed both ON culture and diluted cultures
  • Heated TSB slightly to dissolve glucose
  • Placed in Incubator besides the shaker (37C, static condition at 5pm), incubate for 24 hours
  • To be taken out for washing and fixation before 5pm August 2nd.