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Team:UNIPV-Pavia/Protocols/Ligation
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 Dry Lab
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The protocols we used
- LB medium preparation
- Plasmid resuspension from IGEM paper spots
- Transformation
- Plasmid extraction
- BioBrick digestion with restriction enzymes
- DNA gel extraction
- Antarctic Phosphatase
- Ligation
- PCR
Ligation
(estimated time: 20 min + 12-16 hours overnight incubation)
Materials needed:
- MgCl2
- Buffer
- dNTPs
- ddH2O
- Taq Polymerase
- VF2 primer
- VR primer
- For every DNA sample you want to amplify, put:
- 2 µl buffer
- 0.6 µl MgCl2
- 0.4 µl dNTPs
- Now you can add a loading buffer to the solution and perform electrophoresis to check the amplified sequence length.
