Team:Paris/August 8
From 2008.igem.org
(Difference between revisions)
(→Digestion of PCR products) |
(→Results) |
||
Line 267: | Line 267: | ||
* Incubate 2h30 at 37°C | * Incubate 2h30 at 37°C | ||
- | ==== | + | ==== Purification/Analysis ==== |
'''Electrophoresis''' | '''Electrophoresis''' | ||
- | * ladder : 10µl ladder 1 kb | + | * ladder : 10µl ladder 1 kb (for 1%) or ladder 100pb ( for 1.5%) |
- | * samples : 3µl of | + | * 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 for the '''vector''' and '''1.5%''' for the '''insert'''. |
* Results : | * Results : | ||
Line 285: | Line 285: | ||
|align="center"|'''Expected size''' | |align="center"|'''Expected size''' | ||
|align="center"|'''Measured size''' | |align="center"|'''Measured size''' | ||
- | |||
|- style="text-align: center;" | |- style="text-align: center;" | ||
|align="center"|D139 | |align="center"|D139 |
Revision as of 13:19, 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 Protocol
For each samples, 1 µl dNTP
PCR verification/AnalyseAfter the PCR :
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
Electrophoresis Purification of PCRWhen the PCR cycles were finished, conditions :
|