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Team:EPF-Lausanne/Protocols
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* [[Team:EPF-Lausanne/Protocols/Competent cells|Competent Cells]]: prepare cells for plasmid uptake. | * [[Team:EPF-Lausanne/Protocols/Competent cells|Competent Cells]]: prepare cells for plasmid uptake. | ||
* [[Team:EPF-Lausanne/Protocols/Competent cells. Protocol II|Competent Cells. Protocol II (Inoue method)]]. | * [[Team:EPF-Lausanne/Protocols/Competent cells. Protocol II|Competent Cells. Protocol II (Inoue method)]]. | ||
| - | * [[Team:EPF-Lausanne/Protocols/Glycerol stock|Glycerol stock]]: stock transformed cells at -80°C | + | * [[Team:EPF-Lausanne/Protocols/Glycerol stock|Glycerol stock]]: stock transformed cells at -80°C. |
| + | * [[Team:EPF-Lausanne/Protocols/Agar Plates|Agar plate preparation]]: preparing agar plates for cell cultures. | ||
=== Cloning, assembly, and mutations === | === Cloning, assembly, and mutations === | ||
Revision as of 15:00, 20 July 2011
Protocols
Contents |
Molecular Biology
Products and stock preparation
- Primer preparation: initial dilution of ordered primers.
- Competent Cells: prepare cells for plasmid uptake.
- Competent Cells. Protocol II (Inoue method).
- Glycerol stock: stock transformed cells at -80°C.
- Agar plate preparation: preparing agar plates for cell cultures.
Cloning, assembly, and mutations
- Transformation: introduce foreign plasmid into competent cells.
- Plating: plate transformed cells
- Liquid cultures: when cells have grown on plates, put them in liquid cultures
- Gibson assembly.
- Linear template- TetR.
- tetR Overlap Extension PCR: induce specific mutations on tetR linear template.
- tetR Site-specific mutagenesis: induce site-specific mutations on a plasmid containing tetR.
- Agarose gel electrophoresis: DNA samples analysis (can be followed by band purification).
DNA recovery
- Plasmid preparation - Miniprep: recover plasmids from a bacterial culture.
- Gel purification: recover DNA separated by electrophoresis
Biochemistry
- IPTG test: test expression of a gene downstream from Plac
Microfluidics
- PDMS two layer device fabrication
- MITOMI: Protein – DNA interactions
- Chemostat cell culture
- Klenow dsDNA synthesis
