Team:Paris/August 17
From 2008.igem.org
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{{Paris/Calendar_Links|August 16|August 18}} | {{Paris/Calendar_Links|August 16|August 18}} | ||
- | == | + | =Construction of pLas-TetR-GFP tripart & rbs-LasR-dble ter= |
+ | ==Analysis of the transformation we did [[Team:Paris/August 16 |yesterday]]== | ||
{|border="1" style="text-align: center" | {|border="1" style="text-align: center" | ||
- | === | + | ===Number of colonies and flurorescence=== |
|'''Ligation name''' | |'''Ligation name''' | ||
|'''Description''' | |'''Description''' | ||
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- | === | + | ===PCR Screening=== |
- | + | [[Team:Paris/Notebook/Protocols#PCR Screening| Protocol]] | |
- | [[Image:Screen L150-L151.2.JPG|thumb|L150 | + | [[Image:Screen L150-L151.2.JPG|thumb|L150 and L151]] |
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L150 doesn't success, measured size match with the promoter size only, we will check the size of L101 by screening and if it doesn't match with the good size (1827pb) we will try again the ligation : rbs-TetR (S03879) with GFP tripart (E0840).<br>L151 is ok we have to put a strong promoter before the part rbs-lasR-double terminator :GREAT ! | L150 doesn't success, measured size match with the promoter size only, we will check the size of L101 by screening and if it doesn't match with the good size (1827pb) we will try again the ligation : rbs-TetR (S03879) with GFP tripart (E0840).<br>L151 is ok we have to put a strong promoter before the part rbs-lasR-double terminator :GREAT ! | ||
- | === | + | ===Minipreps=== |
{|border="1" style="text-align: center" | {|border="1" style="text-align: center" | ||
|'''Miniprep Name''' | |'''Miniprep Name''' |
Revision as of 13:24, 30 August 2008
Construction of pLas-TetR-GFP tripart & rbs-LasR-dble terAnalysis of the transformation we did yesterday
PCR Screening
Conclusion : L150 doesn't success, measured size match with the promoter size only, we will check the size of L101 by screening and if it doesn't match with the good size (1827pb) we will try again the ligation : rbs-TetR (S03879) with GFP tripart (E0840). MiniprepsAnalysis 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 ScreeningMiniprepsConstruction of OmpR*+RBS and EnvZ*+RBS: transformationEvaluation of the number of colonies
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