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Team:Lyon-INSA-ENS/Realisation/Week12
From 2011.igem.org
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| - | TSS transformation of MC4100 with pIG27 (Cm) or pIG16 (Amp) | + | <b>TSS transformation</b> of MC4100 with pIG27 (Cm) or pIG16 (Amp) |
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| - | E+P digestion and electrophoresis of all the isolated clones for the following transformations : MC4100+pIG27, MC4100 + pIG16, MC4100 + rcn-ompR234.<br/> | + | E+P <b>digestion and electrophoresis</b> of all the isolated clones for the following transformations : MC4100+pIG27, MC4100 + pIG16, MC4100 + rcn-ompR234.<br/> |
| - | All clones are correct for pIG27 and pIG16. No clone is correct for rcn-ompR234 | + | All clones are correct for pIG27 and pIG16. No clone is correct for rcn-ompR234 -> detection of an error in the ligation. <br/> |
Storage of clone 1 for pIG27 and pIG16. Plating of those same clones on LB+antibiotic (Cm and Amp respectively ).<br/> | Storage of clone 1 for pIG27 and pIG16. Plating of those same clones on LB+antibiotic (Cm and Amp respectively ).<br/> | ||
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| - | Nanodrop quantification of the previous pIG27 miniprep : 48.0 ng/µL.<br/> | + | <b>Nanodrop quantification</b> of the previous pIG27 miniprep : 48.0 ng/µL.<br/> |
Preparation of tubes for sequencing (30µL at approximately 50-100 ng/µL). The pIG27 miniprep is used as such. pIG25 is diluted from 5µL of the collection (371.4 ng/µL) with 25µL water. | Preparation of tubes for sequencing (30µL at approximately 50-100 ng/µL). The pIG27 miniprep is used as such. pIG25 is diluted from 5µL of the collection (371.4 ng/µL) with 25µL water. | ||
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Revision as of 13:23, 13 September 2011
Week 12
From Monday the 5th of September to Friday the 9th of September 2011
Monday
Methyl violet coloration of the flocculation tests from the previous week.
Start of 3 replica of 24 well plates containing :
MC4100+pIG6, MC4100+pIG6+10µM Co ( controls )
MC4100+pIG2 with increasing concentration in CoCl2 (0, 10, 25, 50, 100 µM )
Revelation of the 2 plates from september 2nd.
Tuesday
TSS transformation of MC4100 with pIG27 (Cm) or pIG16 (Amp)
Wednesday
Results of tuesday transformations : no clone on the negative controls, several on the tests. Isolation of 4 clones per transformation in solid and liquid culture.
Pouring of Amp plates.
Thursday
E+P digestion and electrophoresis of all the isolated clones for the following transformations : MC4100+pIG27, MC4100 + pIG16, MC4100 + rcn-ompR234.
All clones are correct for pIG27 and pIG16. No clone is correct for rcn-ompR234 -> detection of an error in the ligation.
Storage of clone 1 for pIG27 and pIG16. Plating of those same clones on LB+antibiotic (Cm and Amp respectively ).
Friday
Nanodrop quantification of the previous pIG27 miniprep : 48.0 ng/µL.
Preparation of tubes for sequencing (30µL at approximately 50-100 ng/µL). The pIG27 miniprep is used as such. pIG25 is diluted from 5µL of the collection (371.4 ng/µL) with 25µL water.
