Team:Paris/August 8
From 2008.igem.org
(→PCR Screening of Ligation Transformants) |
(→Protocol of screening PCR) |
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- | * 50µl of Mix | + | * 50µl of PCR Mix by tube/clone |
- | * one toothpick of each clone's colony | + | * one toothpick of each clone's colony per tube |
* Program : Annealing 55°C - Time élongation 1'30" - Number cycle : 29 | * Program : Annealing 55°C - Time élongation 1'30" - Number cycle : 29 | ||
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=== Conditions of electrophoresis === | === Conditions of electrophoresis === |
Revision as of 14:58, 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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