Team:Evry/Notebook/Transformation/09-08-2014

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<u> <b> Transformation of Pseudovrbrio with pBBR1MCS and pRhokHI-2 - Confirmation </b> </u> <br>
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<u> <b> <big><FONT COLOR=#003333>Transformation of Pseudovrbrio with pBBR1MCS and pRhokHI-2 - Confirmation</font></big> </b> </u> <br>
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We made a restriction control of our plasmid, and we tried to prove that these plasmids are present in our Pseudovibrio. <br>
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<p>We made a restriction control of our plasmid, and we tried to prove that these plasmids are present in our Pseudovibrio. <br>
After made a purification of plasmids in A1 and A2 with NucleoSpin Plamsid Kit (Macherey Nagel), we obtained concentrations at 380.1 ng/µL for pRhokHI-2 and at 91.1 ng/µL for pBBR1MCS.<br> <br>
After made a purification of plasmids in A1 and A2 with NucleoSpin Plamsid Kit (Macherey Nagel), we obtained concentrations at 380.1 ng/µL for pRhokHI-2 and at 91.1 ng/µL for pBBR1MCS.<br> <br>
We disgested these sample and our stock of pRhokHI-2 and pBBR1MCS with XbaI (unique restriction site on these plasmids) and we have make migrate the product of digestion. <br>
We disgested these sample and our stock of pRhokHI-2 and pBBR1MCS with XbaI (unique restriction site on these plasmids) and we have make migrate the product of digestion. <br>
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There was a problem with the digestion. We are going to try with another enzyme.
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There was a problem with the digestion. We are going to try with a longer time of incubation.
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<u> <b> Stock of pBBR1MCS and pRhokHI-2 </b> </u> <br>
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To remake a stock of these plasmids, we put in liquid LB+Cam (1:1000) culture, a colony from a LB+Cam plate of E.Coli transformed.
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Latest revision as of 22:00, 17 October 2014

Picture

Transformation of Pseudovrbrio with pBBR1MCS and pRhokHI-2 - Confirmation

We made a restriction control of our plasmid, and we tried to prove that these plasmids are present in our Pseudovibrio.
After made a purification of plasmids in A1 and A2 with NucleoSpin Plamsid Kit (Macherey Nagel), we obtained concentrations at 380.1 ng/µL for pRhokHI-2 and at 91.1 ng/µL for pBBR1MCS.

We disgested these sample and our stock of pRhokHI-2 and pBBR1MCS with XbaI (unique restriction site on these plasmids) and we have make migrate the product of digestion.

IMAGE
Gel of digestion with XbaI

There was a problem with the digestion. We are going to try with a longer time of incubation.

Stock of pBBR1MCS and pRhokHI-2
To remake a stock of these plasmids, we put in liquid LB+Cam (1:1000) culture, a colony from a LB+Cam plate of E.Coli transformed.

Sep 08