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Team:Lyon-INSA-ENS/Realisation/Week5
From 2011.igem.org
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Start of 5mL cultures of 18A (Amp), 24E (Amp) and Curli(Cn). Incubation at 37°C <br/> | Start of 5mL cultures of 18A (Amp), 24E (Amp) and Curli(Cn). Incubation at 37°C <br/> | ||
| - | Visit of Centraco | + | Visit of Centraco : a french nuclear waste treatment plant <br/> |
Start of 50 mL cultures ( 50mL sterile LB, 500µL antibiotic, 100µL from the 5mL cultures ). Incubation at 37°C overnight. | Start of 50 mL cultures ( 50mL sterile LB, 500µL antibiotic, 100µL from the 5mL cultures ). Incubation at 37°C overnight. | ||
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| - | Miniprep of the ligations from the previous week to check the insert : 5L+2L, 5L+24E, 2M+2L, 2M+24E, 2L-Tet, 24E-Tet<br/> | + | <b>Miniprep</b> of the ligations from the previous week to check the insert : 5L+2L, 5L+24E, 2M+2L, 2M+24E, 2L-Tet, 24E-Tet<br/> |
4 clones are chosen for each ligation. <br/> | 4 clones are chosen for each ligation. <br/> | ||
| - | Digestion and electrophoresis : no clone seemed to contain the insert. Decision to verify the digestion, ligation and transformation protocols. <br/><br/> | + | <b>Digestion</b> and <b>electrophoresis</b> : no clone seemed to contain the insert. Decision to verify the digestion, ligation and transformation protocols. <br/><br/> |
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| - | Midiprep of 18A, 24E and curli parts.<br/> | + | <b>Midiprep</b> of 18A, 24E and curli parts.<br/> |
Nanodrop analysis : <br/> | Nanodrop analysis : <br/> | ||
-18A : 16.7 ng/µL <br/> | -18A : 16.7 ng/µL <br/> | ||
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Start of 50mL cultures for the same parts.<br/><br/> | Start of 50mL cultures for the same parts.<br/><br/> | ||
| - | Miniprep of the clones with the mutated plasmid to get more plasmid for the next step. <br/><br/> | + | <b>Miniprep</b> of the clones with the mutated plasmid to get more plasmid for the next step. <br/><br/> |
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French national day but... we worked.<br/><br/> | French national day but... we worked.<br/><br/> | ||
| - | Midiprep of 22M, 2M, 2L, 5L, OmpR234, RBS-GFP-LVA. | + | <b>Midiprep</b> of 22M, 2M, 2L, 5L, OmpR234, RBS-GFP-LVA. |
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Revision as of 13:00, 13 September 2011
Week 5
From Monday the 11th of July to Friday the 15th of July 2011
Monday
Start of 5mL cultures of 18A (Amp), 24E (Amp) and Curli(Cn). Incubation at 37°C
Visit of Centraco : a french nuclear waste treatment plant
Start of 50 mL cultures ( 50mL sterile LB, 500µL antibiotic, 100µL from the 5mL cultures ). Incubation at 37°C overnight.
Tuesday
Miniprep of the ligations from the previous week to check the insert : 5L+2L, 5L+24E, 2M+2L, 2M+24E, 2L-Tet, 24E-Tet
4 clones are chosen for each ligation.
Digestion and electrophoresis : no clone seemed to contain the insert. Decision to verify the digestion, ligation and transformation protocols.
Midiprep of 18A, 24E and curli parts.
Nanodrop analysis :
-18A : 16.7 ng/µL
-Curli : 40.5 ng/µL
-24E : 190.8 ng/µL
The sequencing results were not enough good due to too high concentration of mutated plasmid. Resent of dillute sample.
Wednesday
Start of 5mL cultures of 22M, 2M, 2L, 5L, OmpR234 and RBS-GFP-LVA ( synthesized part with an unstable GFP )
Start of 50mL cultures for the same parts.
Miniprep of the clones with the mutated plasmid to get more plasmid for the next step.
Another french newspaper "Le progrès" interviewed us.
Thursday
French national day but... we worked.
Midiprep of 22M, 2M, 2L, 5L, OmpR234, RBS-GFP-LVA.
