Team:Paris/August 8
From 2008.igem.org
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* ladder : 10µl ladder 1 kb | * ladder : 10µl ladder 1 kb | ||
* samples : 3µl of insert + 2µl of Loading Dye | * samples : 3µl of insert + 2µl of Loading Dye | ||
- | * Conditions : migration 30min at 100W, on a '''1''' agarose gel. | + | * Conditions : migration 30min at 100W, on a '''1%''' agarose gel. |
* Results : | * Results : | ||
Line 287: | Line 287: | ||
|D139 | |D139 | ||
|FlhDC gene | |FlhDC gene | ||
- | | | + | |4 |
- | |style="background: # | + | |972 pb |
- | + | |style="background: #ff6d73"|<center> 0 pb</center> | |
|- style="text-align: center;" | |- style="text-align: center;" | ||
|D140 | |D140 | ||
|OmpR gene | |OmpR gene | ||
- | | | + | |5 |
- | |style="background: #cbff7B"|<center> | + | |762 pb |
- | + | |style="background: #cbff7B"|<center> 700 pb</center> | |
|- style="text-align: center;" | |- style="text-align: center;" | ||
|D141 | |D141 | ||
|EnvZ gene | |EnvZ gene | ||
- | | | + | |6 |
- | |style="background: #cbff7B"|<center> | + | |1421 pb |
- | + | |style="background: #cbff7B"|<center> 1000 pb</center> | |
|- style="text-align: center;" | |- style="text-align: center;" | ||
|D142 | |D142 | ||
- | + | | -vector digested- | |
- | | | + | |7 |
- | | | + | |2057 pb |
- | |style="background: #cbff7B"|<center> | + | |style="background: #cbff7B"|<center> 2000 pb</center> |
- | | | + | |- style="text-align: center;" |
+ | |D142 | ||
+ | | -vector not digested- | ||
+ | |8 | ||
+ | |2764 pb | ||
+ | |style="background: #ff6d73"|<center> 2100 pb</center> | ||
|} | |} | ||
Revision as of 14:54, 11 August 2008
Minipreps : Plasmid extractionExtraction of pSB3K3 et E0240 in pSB1A2 plasmid from overnight bacteria culture using the QIAspin Miniprep Kit (QIAGEN) by QIACube.
Amplification of Genes of interest (OmpR, EnvZ, FlhDC)We performed PCR on to amplify the sequence in order to have enough amount of DNA to carry out the following of our experiments.
PCR amplificationProtocol
For each samples, 1 µl dNTP
PCR verification/AnalyseAfter the PCR :
==> Conclusion : we observed the size expected for the PCR products, but not for FlhDC gene.
Digestion of PCR productsProtocol :
Purification/AnalysisElectrophoresis
Transformation results
PCR Screening of Ligation TransformantsUse of 8 clones of Ligation transformants for screening PCR
Protocol of screening PCR
Conditions of electrophoresis
Results of electrophoresis
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