Team:Paris/August 8
From 2008.igem.org
(→Purification/Analysis) |
(→PCR Screening of Ligation Transformants) |
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== '''PCR Screening of Ligation Transformants'''== | == '''PCR Screening of Ligation Transformants'''== | ||
- | Use of 8 clones of Ligation transformants for | + | Use of 8 clones of Ligation transformants for PCR screening |
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| rowspan="8" align="center"|L131 | | rowspan="8" align="center"|L131 | ||
- | | rowspan="8" align="center"| | + | | rowspan="8" align="center"|pFlhDC |
|align="center"|1 | |align="center"|1 | ||
|align="center"|red | |align="center"|red | ||
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|align="center"|2X | |align="center"|2X | ||
|- | |- | ||
- | |align="center"| | + | |align="center"|forward primer |
|align="center"|1µl | |align="center"|1µl | ||
|align="center"|10µM | |align="center"|10µM | ||
|- | |- | ||
- | |align="center"| | + | |align="center"|reverse primer |
|align="center"|1µl | |align="center"|1µl | ||
|align="center"|10µM | |align="center"|10µM |
Revision as of 14:57, 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 PCR screening
Protocol of screening PCR
Conditions of electrophoresis
Results of electrophoresis
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