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Team:Evry/Notebook/Transformation/09-08-2014
From 2014.igem.org
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| - | <u> <b> Transformation of Pseudovrbrio with pBBR1MCS and pRhokHI-2 - Confirmation </b> </u> <br> | + | <u> <b> <big><FONT COLOR=#003333>Transformation of Pseudovrbrio with pBBR1MCS and pRhokHI-2 - Confirmation</font></big> </b> </u> <br> |
| - | We made a restriction control of our plasmid, and we tried to prove that these plasmids are present in our Pseudovibrio. <br> | + | <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 | + | 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> | ||
| + | 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. | ||
</p> | </p> | ||
Latest revision as of 22:00, 17 October 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.
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.
