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Team:UNIPV-Pavia/Protocols/Resuspension
From 2008.igem.org
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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
- DNA precipitation with sodium acetate
- Antarctic Phosphatase
- Ligation
- PCR
Plasmid resuspension from IGEM paper spots
(estimated time: 25 min + 5 min for every part if you use scalpel/tweezers or + 15 min for every part if you use punch tool)
Materials needed:
- Pre-warmed at 42°C TE
- Desired spot location information
- Scalpel and tweezers (or punch tool)
- ddH20
- 99% ethanol
- 0.5 ml tubes
- Put 10 µl of pre-warmed TE into a 0.5 ml tube.
- Cut paper spots using scalpel and tweezers (or punch tool, following the instructions provided with the IGEM kit); if you use scalpel and tweezers, try to cut pieces of about the same dimension of the punch tool.
- Put the cut paper into the 0.5 ml tube.
- Clean scalpel and tweezers (or punch tool) with water and ethanol every time you cut a spot; be careful to dry your tools correctly, especially if you use punch tool, which needs much more time to dry than scalpel/tweezers.
- Incubate at 42°C for 20 min.
- Vortex and spin down.
