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

{| width="800"
 * style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]] iGEM Project name 1
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

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 Column Strain Strain:TSB Glucose % (w/v) A JG 1:100 10 B JG 1:100 20 D MW 1:50 20 E MW 1:50 10 F Control - 10 G Control - 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.


 * }