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Team:Cambridge/Lab book/Week 8
From 2012.igem.org
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*Verified with nanodropper - final DNA concentrations: | *Verified with nanodropper - final DNA concentrations: | ||
| + | |||
| + | '''[[Team:Cambridge/Protocols/beta-galactosidaseassay| Characterization of fluoride riboswitch construct]]''' | ||
| + | |||
| + | ---- | ||
| + | |||
| + | *Strain of ''bacillus'' lacking fluoride transporter system tested. Concentrations used: | ||
| + | |||
| + | :*0mM | ||
| + | :*0.5mM | ||
| + | :*1mM | ||
| + | :*2.5mM | ||
| + | :*5mM | ||
| + | :*10mM | ||
| + | :*20mM | ||
| + | :*30mM | ||
===Thursday (16/08/12)=== | ===Thursday (16/08/12)=== | ||
Revision as of 17:45, 15 August 2012
| Week: | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
|---|
Contents |
Monday (13/08/12)
Tuesday (14/08/12)
Gibson assembly of positive control
- Fragments used:
- Protocol changed slightly: 0.5 μl of each DNA fragment solution mixed in with master mix to make up 4 μl total volume (1μl DNA and 3μl master mix).
Transformation of e.coli with positive control DNA
- Chemically competent e.coli cells transformed with plasmid DNA produced by Gibson assembly step.
- Transformants plated out on 50 μg/ml kanomycin plates. Incubated overnight at 37 °C.
Wednesday (15/08/12)
PCR of positive control fragments
- Standard PCR settings used
Extraction of positive control DNA
- Positive control DNA from gel excised and purified.
- Additional elution steps used to concentrate resultant solution.
- Verified with nanodropper - final DNA concentrations:
Characterization of fluoride riboswitch construct
- Strain of bacillus lacking fluoride transporter system tested. Concentrations used:
- 0mM
- 0.5mM
- 1mM
- 2.5mM
- 5mM
- 10mM
- 20mM
- 30mM
Thursday (16/08/12)
Friday (17/08/12)
