Difference between revisions of "Team:Aix-Marseille/Notebook"
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* Reisolement of the knockout fliC mutant W3110 from the transformation of the 09.08.2016 | * Reisolement of the knockout fliC mutant W3110 from the transformation of the 09.08.2016 | ||
* Miniprep of the BBa_K1951008 + BbsI 215-216 position and starters of BBa_K1951008 + BbsI 238-239 position | * Miniprep of the BBa_K1951008 + BbsI 215-216 position and starters of BBa_K1951008 + BbsI 238-239 position | ||
| − | * Digestion test by BBs1 and EcoRI using NEB buffer and electrophoresis | + | * Digestion test by BBs1 and EcoRI using NEB buffer and electrophoresis analyse |
| − | * Colony PCR of BBa_K1951011 in pSB1C3 by | + | * Colony PCR of BBa_K1951011 in pSB1C3 by amplification with different SLIC oligos FW and RV to see the differente constitutive biobrick. Biobrick is OK. |
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===== Satursday, September 10th ===== | ===== Satursday, September 10th ===== | ||
| − | * Swimming test of the knockout fliC mutant W3110 and the WT | + | * Swimming test of the knockout fliC mutant W3110 for 10 differents colonies and the W3110 WT as a positive temoin. |
* Starters of the fliC mutant W3110 | * Starters of the fliC mutant W3110 | ||
| + | * Monitoring growth from differents strain to make comassie. From starters of biobrick BBa_K1951008, BBa_K1951009, BBa_K1951010, Tg1 WT (negative temoin), Tg1+pSB1C3 containing RFP (negative temoin), and BBa_K1951011, we made cultures at DO=0.2 in duplicate. At DO=0.6, half of the cultures has been induced with 0.02 arabinose for the biobrick containing an inductible promotor. Every hour, a sample of 1uDO was taken. After centrigugation at 5000g during 5min and removing of the supernatant, we added 50µL beta mercatoethanol and congeled at -20°C to test protein production on a SDS page/Comassie later. | ||
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===== Sunday, September 11st ===== | ===== Sunday, September 11st ===== | ||
| − | * Made competant of the fliC mutant W3110 strain | + | * Made competant of the fliC mutant W3110 strain from 5 differents colonies from 4 colonies which swam well |
| − | * Transformation of the previous competant with BBa_K1951008 | + | * Transformation of the previous competant with BBa_K1951008 and BBa_K1951009 |
| + | * Transformation of cadA mutant Keio strain with BBa_K1951011 | ||
</div> | </div> | ||
| − | <div id="2016-09- | + | <div id="2016-09-12" class="evday"> |
| − | ===== | + | ===== Monday, September 12rd ===== |
| + | |||
| + | * Transformation of the fliC mutant W3110 competant with BBa_K1951008 | ||
| + | |||
| + | </div> | ||
| + | <div id="2016-09-13" class="evday"> | ||
| + | |||
| + | ===== Tuesday, September 13th ===== | ||
| + | |||
| + | * Swimming test of colonies obtained after transformation of the 09.12.2016. After 3h incubation at 37°C, swimming has been recovered | ||
| + | |||
| + | |||
| + | </div> | ||
| + | <div id="2016-09-15" class="evday"> | ||
| + | |||
| + | ===== Thursday, September 15===== | ||
| − | * | + | * Dillution of peptides oligos by 10000 (ex: 0.31mg/238µL/10000=ng/µL |
| − | * | + | * Annealing 98°C during 2min and cool down during approximatly 3h |
| + | * Ligation of 5ng from the annealing mix for 100ng pSB1C3 digested by EcoRI and PstI | ||
| + | * Transformation in Tg1 | ||
</div> | </div> | ||
{{:Team:Aix-Marseille/Template-Footer}} | {{:Team:Aix-Marseille/Template-Footer}} | ||
