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Team:EPF Lausanne/SecondWeek
From 2010.igem.org
(Difference between revisions)
| Line 27: | Line 27: | ||
*Measure OD of overnight culture of Asaia for competence --> value was too high | *Measure OD of overnight culture of Asaia for competence --> value was too high | ||
*Diluted culture to 0.3 OD, left in 30 degree incubator for 2 hours, OD of 0.6 measured --> start with competence | *Diluted culture to 0.3 OD, left in 30 degree incubator for 2 hours, OD of 0.6 measured --> start with competence | ||
| - | *Asaia competence | + | *Asaia competence |
*Digestion of BB's produced and of the Base vector | *Digestion of BB's produced and of the Base vector | ||
| - | *Agar plates with Kan/Tet/Cm made | + | *Agar plates with Kan/Tet/Cm/Amp made |
| + | *Run Gel with the products of the digestion to check if the fragments are as expected | ||
| + | *Prepare Tet stock (from powder) | ||
<h3>Thursday 22.07.2010 </h3> | <h3>Thursday 22.07.2010 </h3> | ||
<h3>Friday 23.07.2010 </h3> | <h3>Friday 23.07.2010 </h3> | ||
Revision as of 16:04, 21 July 2010
Contents |
Monday 19.07.2010
- Results of the transformation checked: successful!!
- Amplification of colonies (goal to do a miniprep of the plasmids --ori-- and --ori-resistance--)
- Results of plating of asaia culture (where we hope to recover the WT) checked: Asaia cultures grew on Kan plates also! Checked colonies for fluorescance under the microscope: no fluorescance --> Very unexpected! Don't know reason yet.
- Re-grow the colonies. Multiple cycles: hope that we get WT.
- Preparation of Asaia cultures for Lemaitre experiments/WT/Competence...
- Preparation of SOC medium and GLY medium
- Autoclaving of media and flasks
Tuesday 20.07.2010
- Plating Asaia to get WT (once again)
- Making HEPES Buffer for the Competence of Asaia
- Diluted the culture to get a 25ml culture(1:20) according to Asaia competence protocol
- OD curve plotted, Doubling time calculated
- No single colonies from the re-growing trial --> replating properly on separate plates
- Preparation of LB medium mixed with Kan/Amp/Cm and GLY medium mixed with <kan
- Miniprep of Origin + Resistance
- Miniprep of Origin alone.
- Miniprep of base vector BBa_151020
- Concentration measurments of miniprep products
- Glycerol stock of BB plasmid and Base Vector made
Wednesday 21.07.2010
- Measure OD of overnight culture of Asaia for competence --> value was too high
- Diluted culture to 0.3 OD, left in 30 degree incubator for 2 hours, OD of 0.6 measured --> start with competence
- Asaia competence
- Digestion of BB's produced and of the Base vector
- Agar plates with Kan/Tet/Cm/Amp made
- Run Gel with the products of the digestion to check if the fragments are as expected
- Prepare Tet stock (from powder)
