"
Team:EPF-Lausanne/Protocols
From 2011.igem.org
(Difference between revisions)
(→Cloning, assembly, and mutations) |
(→Products and stock preparation) |
||
| Line 7: | Line 7: | ||
* [[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 | ||
=== Cloning, assembly, and mutations === | === Cloning, assembly, and mutations === | ||
Revision as of 08:10, 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.
- 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
