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Team:UNIPV-Pavia/Methods Materials/Transformation
From 2009.igem.org
(Difference between revisions)
(New page: {{UNIPV-Pavia/Protocols}} <br> == '''Transformation''' == ''(estimated time: 3 hours and 30 min + 12-16 hours overnight incubation)'' <br> <br> '''Materials needed:''' *'''LB agar plates ...) |
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*'''SOC medium''' | *'''SOC medium''' | ||
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| - | *Put | + | *Put 1-2 µl of DNA resuspension or ligation into TOP10 tube. |
| - | *Incubate on ice for 30 | + | *Incubate on ice for 30 min. |
*Heat shock: 42°C for 1 min. | *Heat shock: 42°C for 1 min. | ||
*Put transformed TOP10 tube on ice and then add 250 µl SOC medium. | *Put transformed TOP10 tube on ice and then add 250 µl SOC medium. | ||
| - | *Incubate | + | *Incubate 1 hour at 37°C, 220 rpm. |
| - | * | + | *Plate 200 µl of the solution on a proper agar plate. |
| - | + | ||
| - | + | ||
*Incubate overnight at 37°C. | *Incubate overnight at 37°C. | ||
<br> | <br> | ||
Revision as of 16:51, 22 June 2009
- LB medium
- DNA resuspension from iGEM 2009 plates
- Transformation
- X-Gal staining protocol for beta galactosidase (blue/white screening)
- Glycerol stocks
- Plasmid extraction
- BioBrick digestion with restriction enzymes
- Electrophoresis
- DNA gel extraction
- DNA precipitation with sodium acetate
- Ligation
- PCR
- Sensing ethanol concentration: our do-it-yourself kit
Transformation
(estimated time: 3 hours and 30 min + 12-16 hours overnight incubation)
Materials needed:
- LB agar plates with proper antibiotic added incubated at 37°C
- Thawed Invitrogen TOP10 cells (every tube contains approximately 60 µl of competent cells)
- Resuspended DNA
- SOC medium
- Put 1-2 µl of DNA resuspension or ligation into TOP10 tube.
- Incubate on ice for 30 min.
- Heat shock: 42°C for 1 min.
- Put transformed TOP10 tube on ice and then add 250 µl SOC medium.
- Incubate 1 hour at 37°C, 220 rpm.
- Plate 200 µl of the solution on a proper agar plate.
- Incubate overnight at 37°C.
