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Team:Evry/Notebook/Transformation/09-08-2014
From 2014.igem.org
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There was a problem with the digestion. We are going to try with a longer time of incubation. | 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> | <u> <b> Stock of pBBR1MCS and pRhokHI-2 </b> </u> <br> | ||
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. | 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. | ||
Revision as of 15:21, 12 September 2014
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.
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
