"
Team:EPF-Lausanne/Protocols
From 2011.igem.org
(Difference between revisions)
(→Products and stock preparation) |
(→Cloning, assembly, and mutations) |
||
| Line 12: | Line 12: | ||
* [[Team:EPF-Lausanne/Protocols/Transformation|Transformation]]: introduce foreign plasmid into competent cells. | * [[Team:EPF-Lausanne/Protocols/Transformation|Transformation]]: introduce foreign plasmid into competent cells. | ||
| + | * [[Team:EPF-Lausanne/Protocols/Plating|Plating]]: plate transformed cells | ||
| + | * [[Team:EPF-Lausanne/Protocols/Liquid cultures|Liquid cultures]]: when cells have grown on plates, put them in liquid cultures | ||
* [[Team:EPF-Lausanne/Protocols/Gibson assembly|Gibson assembly]]. | * [[Team:EPF-Lausanne/Protocols/Gibson assembly|Gibson assembly]]. | ||
* [[Team:EPF-Lausanne/Protocols/TetR|Linear template- TetR]]. | * [[Team:EPF-Lausanne/Protocols/TetR|Linear template- TetR]]. | ||
Revision as of 14:54, 14 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
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.
- 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
Microfluidics
- PDMS two layer device fabrication
- MITOMI: Protein – DNA interactions
- Chemostat cell culture
- Klenow dsDNA synthesis
