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Team:Paris/August 17
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| - | L150 doesn't success, measured size match with the promoter size only, so we think the ligation L101 doesn't succes either. We will try again the ligation : rbs-TetR (S03879) with GFP tripart (E0840). | + | L150 doesn't success, measured size match with the promoter size only, so we think the ligation L101 doesn't succes either. We will try again the ligation : rbs-TetR (S03879) with GFP tripart (E0840).L151 is ok we have to put a strong promoter before the part rbs-lasR-double terminator :GREAT ! |
| - | L151 is ok we have to put a strong promoter before the part rbs-lasR-double terminator :GREAT ! | + | |
==='''Minipreps'''=== | ==='''Minipreps'''=== | ||
Revision as of 16:33, 17 August 2008
Analysis of the transformation we did yesterday
PCR Screening==> Protocol 
L150 doesn't success, measured size match with the promoter size only, so we think the ligation L101 doesn't succes either. We will try again the ligation : rbs-TetR (S03879) with GFP tripart (E0840).L151 is ok we have to put a strong promoter before the part rbs-lasR-double terminator :GREAT ! Minipreps
Analysis of the other transformation we did yesterdayEvaluation of the number of colonies
Remarks:
L143 and L144 did not work once again. Maybe there is a problem with the digestion, because the primers used present only
two nucleotides after the restriction sites. what we should do is cutting pfliL with xbaI and SpeI. As the vector will
autoligate, we should use a phosphatase to prevent this phenomenon.
For L147 and its control (T4), we do not observe any red fluorescence.
It means that the digestion work well, because if it was not digested, the strong promoter J23100 should express mRFP!
We will do the screening anyway.
PCR ScreeningMiniprepsKok-Phen transformation we did yesterdayEvaluation of the number of colonies
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