"
Team:Cambridge/Lab book/Week 8
From 2012.igem.org
(Difference between revisions)
(→Tuesday (14/08/12)) |
(→Tuesday (14/08/12)) |
||
| Line 20: | Line 20: | ||
---- | ---- | ||
| - | *Protocol changed slightly: 0.5 μl of each DNA fragment solution mixed in with master mix to make up 4 μl total volume ( | + | *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). | ||
'''[[Team:Cambridge/Protocols/TransformationofE.coli|Transformation of e.coli with positive control DNA]]''' | '''[[Team:Cambridge/Protocols/TransformationofE.coli|Transformation of e.coli with positive control DNA]]''' | ||
Revision as of 16:59, 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:
Thursday (16/08/12)
Friday (17/08/12)
