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Team:Evry/Notebook/Transformation/08-25-2014
From 2014.igem.org
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<u> <b> Amplification of pRhokHI-2 in E.Coli Bl21 </b> </u> <br> | <u> <b> Amplification of pRhokHI-2 in E.Coli Bl21 </b> </u> <br> | ||
| - | We re-tried to transform E.Coli with pRhokHI-2 to amplify it | + | 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> | <u> <b> Transformation of Pseudovribrio with pBBR1MCS </b> </u> <br> | ||
| - | After made a purification of pBBR1MCS with NucleoSpin Plamsid Kit (Macherey Nagel), electrocompetent Pseudovibrio cells were tranformed with the plasmid by electroporation | + | 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. |
Revision as of 15:12, 2 September 2014
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.
