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Team:EPF-Lausanne/Protocols
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* [[Team:EPF-Lausanne/Protocols/Site-specific mutagenesis|tetR Site-specific mutagenesis]]: induce site-specific mutations on a plasmid containing tetR. | * [[Team:EPF-Lausanne/Protocols/Site-specific mutagenesis|tetR Site-specific mutagenesis]]: induce site-specific mutations on a plasmid containing tetR. | ||
* [[Team:EPF-Lausanne/Protocols/Gel_Electrophoresis|Agarose gel electrophoresis]]: DNA samples analysis (can be followed by band purification). | * [[Team:EPF-Lausanne/Protocols/Gel_Electrophoresis|Agarose gel electrophoresis]]: DNA samples analysis (can be followed by band purification). | ||
| + | * [[Team:EPF-Lausanne/Protocols/Colony PCR|Colony PCR]]: Analyze the plated transformed cells, to see if Gibson assembly worked | ||
=== DNA recovery === | === DNA recovery === | ||
Revision as of 13:33, 3 August 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.
- Autoclave: to sterilize solutions and glassware.
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).
- Colony PCR: Analyze the plated transformed cells, to see if Gibson assembly worked
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
