Team:Evry/Notebook/Transformation/08-25-2014

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<u> <b> Amplification of pRhokHI-2 </b> </u> <br>
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<u> <b> <big><FONT COLOR=#003333>Amplification of pRhokHI-2 in E.Coli Bl21 </font></big></b> </u> <br>
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We re-tried to transform E.Coli with pRhokHI-2 to amplify it <b>(LIEN PROTO)</b>. This time, after a liquid culture of only one hours cells were plated on LB 1X + Kan (1:1000) and let in incubator overnight.
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We re-tried to transform E.Coli with pRhokHI-2 to amplify it (1µL of plasmid for 40µL of cells, 1800V). This time, after a liquid culture of only one hours cells were plated on LB 1X + Kan (1:1000) and let in incubator overnight.
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<u> <b> Transformation of Pseudovribrio with pBBR1MCS </b> </u> <br>
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<u> <b> <big><FONT COLOR=#003333>Transformation of Pseudovribrio with pBBR1MCS</font></big> </b> </u> <br>
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After made a MiniPrep purification of pBBR1MCS <b>(LIEN PROTO)</b>, electrocompetent Pseudovibrio cells were tranformed with the plasmid by electroporation <b>(LIEN PROTO)</b>, then they were incubated for three hours and showed on MB 1X+Cam (1:1000) plates. Plates are let in incubator overnight.
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After made a purification of pBBR1MCS with NucleoSpin Plamsid Kit (Macherey Nagel), electrocompetent Pseudovibrio cells were tranformed with the plasmid by electroporation (1µL of plasmid for 40µL of cells, 1800V), then they were incubated for three hours and showed on MB 1X+Cam (1:1000) plates. Plates are let in incubator overnight.

Latest revision as of 16:55, 12 October 2014

Picture

Amplification of pRhokHI-2 in E.Coli Bl21
We re-tried to transform E.Coli with pRhokHI-2 to amplify it (1µL of plasmid for 40µL of cells, 1800V). This time, after a liquid culture of only one hours cells were plated on LB 1X + Kan (1:1000) and let in incubator overnight.

Transformation of Pseudovribrio with pBBR1MCS
After made a purification of pBBR1MCS with NucleoSpin Plamsid Kit (Macherey Nagel), electrocompetent Pseudovibrio cells were tranformed with the plasmid by electroporation (1µL of plasmid for 40µL of cells, 1800V), then they were incubated for three hours and showed on MB 1X+Cam (1:1000) plates. Plates are let in incubator overnight.

Aug 25