IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/02

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BioFilm Track

  • Made 200ml PBS buffer solution for washing biofilm
  • Leaving 96 well plates with staph strain RN4220 for 48hr incubation - Staph strain RN4220 most likely would not form biofilm according to literature references


Started "tube method" to test staph strain 8325 for biofilm formation

  • Time placed in incubator; 4pm (20 hours incubation, 37C, static condition)


Content

  • 10ml TSB in glass tube
  • 5 % w/v glucuose (0.5g)
  • 2 % w/v NaCl (To facilitate biofilm growth)
  • Staph strain 8325 obtained from Lando

(Heated TSB to dissolve glucose, NaCl. Cooled before adding staph)

dspB Track

Gel Verify

Gel Verify PCR products from last day
Protocol: gel verification protocol in Protocol binder (SOP)

  • Changes: Used 1% agarose gel instead of 0.8%

Gel orientation:

Gel orientation
ABCDEF100bp ladderGHIJKLM

Machine conditions: 100V, 45 min, 0.5X TBE Buffer
Results:
File:100802 DspB 10.tif
Vicki Ma 02:40, 4 August 2010 (EDT)

Restriction Digest(ON)

Protocol: Restriction digest protocol in Protocol binder (SOP)

Table 4. Restriction Digest Supermix
REAGENTS1 RXN VOLUME (uL)SUPERMIX
Buffer 25x630
BSA0.5x63
ddH2O37x6222
EcoRI1x66
PstI1x66
  • DNA: Add 5uL of each A,B,C,D,E,F, psb1A3 (backbone plasmid) [a,b,c,d,e,f]
  • Left ON in 4C fridge on ice

Vicki Ma 03:08, 3 August 2010 (EDT)

Misc

Poured 2* 400ml Amp plate + 1 * 400ml Chloro plate