Team:UNIPV-Pavia/Protocols/Lb
From 2008.igem.org
(Difference between revisions)
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*[[Team:UNIPV-Pavia/Protocols/Digestion|BioBrick digestion with restriction enzymes]] | *[[Team:UNIPV-Pavia/Protocols/Digestion|BioBrick digestion with restriction enzymes]] | ||
*[[Team:UNIPV-Pavia/Protocols/GelExtraction|DNA gel extraction]] | *[[Team:UNIPV-Pavia/Protocols/GelExtraction|DNA gel extraction]] | ||
+ | *[[Team:UNIPV-Pavia/Protocols/Precipitation|DNA precipitation with sodium acetate]] | ||
*[[Team:UNIPV-Pavia/Protocols/AntarcticPhosphatase|Antarctic Phosphatase]] | *[[Team:UNIPV-Pavia/Protocols/AntarcticPhosphatase|Antarctic Phosphatase]] | ||
*[[Team:UNIPV-Pavia/Protocols/Ligation|Ligation]] | *[[Team:UNIPV-Pavia/Protocols/Ligation|Ligation]] |
Latest revision as of 12:51, 1 July 2008
Home | The Team | The Project | Biological Safety | Parts Submitted to the Registry |
---|---|---|---|---|
Dry Lab | Wet Lab | Modeling | Protocols | Activity Notebook |
The protocols we used
- LB medium preparation
- Plasmid resuspension from IGEM paper spots
- Transformation
- Plasmid extraction
- BioBrick digestion with restriction enzymes
- DNA gel extraction
- DNA precipitation with sodium acetate
- Antarctic Phosphatase
- Ligation
- PCR
LB medium preparation
(Estimated time: 10 min + 4 hours of autoclavation and cooling)
Materials needed:
- NaCl
- BactoTryptone
- Yeast extract
- ddH2O
- Agar
- Clean bottle or E-flask
- For a final volume of 1 l, put:
- 10 g NaCl
- 10 g BactoTryptone
- 5 g Yeast extract
- 15 g Agar (only for LB plates)
- into 1 l of ddH2O.
- Autoclave the solution.
- Let it cool to ~40-45°C
- Add the proper antibiotic if needed.
- ONLY FOR PLATES:
- Pour an homogenous layer of agar LB into Petri plates under the hood.
- Let agar LB polymerase.
- Cover and store at +4°C.
- Always check for contaminations before using!