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
1 | 2 | 1kb ladder | 3 | 4 |
Results:
Transformation
- Testing competency
- Transform in pBAD (Amp-resistant)
- Changes: add 1uL of pBAD to competent cells
- Transforming ligation mixes (1,3,4 - all Amp-resistant)
- 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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