Team:Evry/Notebook/Biology/08-15-2014

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      <h2>Title</h2>
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      <p align="justify">  BBa_E0240 and BBa_I20260 are respectively into a PSB1C3 and a PSB1K3 vector. In order to select colonies, LB complemented with kanamycin or chloramphenicol are necessary. 200 ml of LB agar medium was prepared with 7 g of LB Agar powder(Sigma) into 200 ml of miliQ water. After complete dissolution, the solution was sterilized in the Tuttmauer 2540ML apparatus, STE 20 minutes at 121°C and EXT+DRY 15 minutes. 50 µl of kanamycin stock solution (25 mg/L) was added to 50 ml of LB agar to pour 2 plates. 150 µl of chloramphenicol stock solution was added in the 150 LB agar remaining ml to pour 6 plates. 
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<br/> The transformation was performed on DH5 alpha ''E. coli'', as followed: </p>
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<li> Remove E. coli competent tubes from -80°C and keep it on ice
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<li> Add 1 µl of template (here solubilized plasmids from the registry distribution kit) and mix gently
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<li> Incubate 10 minutes on ice
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<li> Perform an heat shock 30 seconds at 42°C
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<li> Incubate 2 minutes on ice
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<li> Add 3 ml of LB medium and incubate 60 minutes at 37°C with an agitation at 200 rpm
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<li> Plate 200 µl of BBa_E0240 on a chloramphenicol LB agar plate and BBa_I20260 on a Knamycin LB agar plate
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<li> Incubate plate overnight at 37°C
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      <span class="cd-date">Aug 15</span>
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Latest revision as of 12:29, 23 August 2014

Retrieved from "http://2014.igem.org/Team:Evry/Notebook/Biology/08-15-2014"