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Théotime
Restriction digest of the same minipreps as yestarday (concentration measured by nanodrop 07/22) using the same protocol as yesterday : 3µg of DNA per reaction.
- RBS/LuxI (K081008) : 32µl DNA + 4µl Buffer 4 10x + 0.4µl BSA 100x + 2µl EcoRI-HF + 2µl SpeI
- TetA(C) (J31007) in a triple volume : 60µl DNA + 7.5µl Buffer 3 10x + 0.8µl BSA 100x + 4µl XbaI + 4µl PstI
- LacZ alpha (I732006) in a triple quantity : : 41µl DNA + 5.5µl Buffer 3 10x + 0.6µl BSA 100x + 4µl XbaI + 4µl PstI
- LuxR (C0062), in triple : 59µl DNA + 7.5µl Buffer 3 10x + 0.7µl BSA 100x + 4µl XbaI + 4µl PstI
- attC : 111µl DNA + 13µl Buffer 2 10x + 1.3µl BSA 100x + 2µl SpeI +2µl PstI
- term/attC/mRFP : 91µl DNA + 11µl Buffer 3 10x + 1.1µl BSA 100x + 2µl XbaI + 2µl PstI
- pLux (R0062) : 49µl DNA + 6µl Buffer 2 10x + 0.6µl BSA 100x + 2µl SpeI + 2µl PstI
- RBS/GFP/term (E0240) : 26µl DNA + 3.5µl Buffer 3 10x + 0.4µl BSA 100x +2 µl XbaI + 2µl PastI
Stéphane
Transformation of TOP10 chemically competent E.coli with the overnight ligations from 07/22. Recuperation for 30 mins after the heat shock in SOC, then plating with 150µl of the transformmation on the ampicilin plate, incubation overnight at 37°C.
- TOP10 pSB1A2 weak RBS -TetA(C)
- TOP10 pSB1A2 med.RBS -TetA(C)
- TOP10 pSB1A2 weak RBS -LuxR
- TOP10 pSB1A2 med.RBS -LuxR
- TOP10 pSB1A2 stand.RBS -LuxR
Théotime and Aleksandra<:font>
Gel electrophoresis of the restriction digest produscts, onmy for the future inserts.
Agarose gel 0.8%, 50V
- TetA(C) (J31007) : 37µl + 7.4µl lb
- TetA(C) (J31007) : 37µl + 7.4µl lb
- -
- LuxR (C0062) : 37µl + 7.4µl lb
- LuxR (C0062) : 37µl + 7.4µl lb
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