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Team:UNIPV-Pavia/Protocols/AntarcticPhosphatase
From 2008.igem.org
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|---|---|---|---|---|
 Dry Lab
|  Wet Lab
|  Modeling
|  Protocols
|  Activity Notebook
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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
Antarctic Phosphatase
(estimated time: 1 hour and 30 min)
Materials needed:
- NEB Antarctic Phosphatase
- 10X NEB Antarctic Phosphatase buffer
- Cut and gel-extracted vector
- Add the proper amount of 10X buffer to a final concentration of 1X (e.g. 2 )
- Cut paper spots using scalpel and tweezers (or punch tool, following provided instructions); 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.
