IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/09

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Phage Track

Eric F.

Prep for UV Induction

Reagents to be prepared

  • Soft Agar - 0.65% Agar
    • Making 400mL, probably only need 26mL tomorrow
    • Recipe: 8.5g LB Agar / 400mL water
  • Hard Agar - 1.5% Agar
    • Making 400mL, probably only need 180mL tomorrow
    • Recipe: 18.5g LB Agar / 400mL water

Autoclaved both at 250 degrees F for 25 minutes

  • Made 1/10 dilution of LB Broth
  • Recipe: 3mL LB Broth + 27mL sdH20


Began overnight cultures of 8325 + RN4220

# Strain Use
1 8325 UV Induction
2 8325 UV Induction
3 RN4220 UV Induction
4 8325 Biofilm Growth
5 RN4220 Biofilm Growth

All overnight cultures were done in 13mL test tubes with 3mL LB Broth. Two overnight controls were used.

Place in incubator @ 37 degrees C, 220 RPM @ 1745.

Tomorrow:

  • 2 independent UV induction protocol run throughs
  • Day 2 of Biofilm protocol
  • Refine Biofilm protocol


dspB Track

Gel verification on ligation mixes from Aug 05 2010

  • Changes: 1% agarose gel
  • Samples: 1,2,3,4
  • Machine conditions: 0.5x TBE buffer, 100V, 45min

Gel orientation:

Gel orientation
121kb ladder34

Results:

Transformation

  1. Testing competency
    1. Transform in pBAD (Amp-resistant)
    2. Changes: add 1uL of pBAD to competent cells
  2. Transforming ligation mixes (1,3,4 - all Amp-resistant)
    1. Changes: add 6uL of each to competent cells

- Both incubate for 1 hour at 37C
- Both spread on Amp plates
- Plates 1,3,4pBAD in 37C at 1540
Vicki Ma 02:55, 11 August 2010 (EDT)



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