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Team:Lyon-INSA-ENS/Realisation/Week10
From 2011.igem.org
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Dissolution of 0.08g of Spectinomycin ( C14H24N2O7 + 2 HCl + 5 H20 ) into 5mL water and filtration to obtain a solution at 10mg/mL.<br/><br/> | Dissolution of 0.08g of Spectinomycin ( C14H24N2O7 + 2 HCl + 5 H20 ) into 5mL water and filtration to obtain a solution at 10mg/mL.<br/><br/> | ||
| - | QIAGen extraction of NM522 and NM522+pIG4 and electrophoresis : no plasmid is in the strains. | + | QIAGen extraction of NM522 and NM522+pIG4 and electrophoresis : no plasmid is in the strains. <br><br> |
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| + | Transformation and plating of NM522 on LB with : pUC18 (=pIG6) (3 µL) + Amp, pIG25 (3 µL) + Cm<br> | ||
| + | Transformation and plating of S19 on LB and the antibiotic Amp with : p157 (2 µL) + SpcR, p115 (2 µL) + SpcR, p127 (2 µL) + Kn, p116 (2 µL) + Kn <br> | ||
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| + | Transformation and plating of NM522 on LB with : p157 (2 µL) + Spc, p115 (2 µL) + Spc, p56 (2 µL) + Amp, p10 (3 µL) + Amp, pIG30 (2 µL) + Cm, p127 (2 µL) + Kn, p116 (2 µL) + Kn <br> | ||
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| + | pIG6 (3µL), pIG7 (3µL), pIG24 (2µL), p56 (2µL), p10 (2µL), pIG16+p157 (3µL each), pIG16+p115(3µL each), pIG16+p127(3µL each), pIG16+p116 ( 3µL and 5µL respectively ), pIG25 (1µL). <br/><br/> | ||
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Revision as of 15:38, 23 August 2011
Week 10
From Monday the 22th of August to Friday the 26th of August 2011
Monday
Both NM522 and NM522/pIG24 grew on LB+Amp ( liquid and solid ) : antibiotic concentration in liquid was not sufficient, a more concentrated solution of Amp (10mg/mL which corresponds to 100X) is made and sterilized by filtration.
Creation of our own LB+Amp plates by plating 200µL of Amp on a LB plate.
Plating of several AmpS strains on the previous plates : PHL818, MC4100, NM522+pIG4, NM522 from the collection, NM522 from the previous LB+Amp plate, a new NM522 found in another lab.
Plating of 10µL of the new NM522 on LB
Dissolution of 0.08g of Spectinomycin ( C14H24N2O7 + 2 HCl + 5 H20 ) into 5mL water and filtration to obtain a solution at 10mg/mL.
QIAGen extraction of NM522 and NM522+pIG4 and electrophoresis : no plasmid is in the strains.
Transformation and plating of NM522 on LB with : pUC18 (=pIG6) (3 µL) + Amp, pIG25 (3 µL) + Cm
Transformation and plating of S19 on LB and the antibiotic Amp with : p157 (2 µL) + SpcR, p115 (2 µL) + SpcR, p127 (2 µL) + Kn, p116 (2 µL) + Kn
Tuesday
Transformation and plating of NM522 on LB with : p157 (2 µL) + Spc, p115 (2 µL) + Spc, p56 (2 µL) + Amp, p10 (3 µL) + Amp, pIG30 (2 µL) + Cm, p127 (2 µL) + Kn, p116 (2 µL) + Kn
pIG6 (3µL), pIG7 (3µL), pIG24 (2µL), p56 (2µL), p10 (2µL), pIG16+p157 (3µL each), pIG16+p115(3µL each), pIG16+p127(3µL each), pIG16+p116 ( 3µL and 5µL respectively ), pIG25 (1µL).
Wednesday
Thursday
Friday
