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Team:UNIPV-Pavia/Protocols/Resuspension
From 2008.igem.org
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<br> | <br> | ||
| + | <h1>The protocols we used</h1> | ||
| + | |||
| + | *[[Team:UNIPV-Pavia/Protocols/Lb|LB medium preparation]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Resuspension|Plasmid resuspension from IGEM paper spots]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Transformation|Transformation]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/PlasmidExtraction|Plasmid extraction]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Digestion|BioBrick digestion with restriction enzymes]] | ||
| + | *[[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/Ligation|Ligation]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Pcr|PCR]] | ||
| + | |||
<h1>Plasmid resuspension from IGEM paper spots</h1> | <h1>Plasmid resuspension from IGEM paper spots</h1> | ||
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*'''Desired spot location information''' | *'''Desired spot location information''' | ||
*'''Scalpel and tweezers (or punch tool)''' | *'''Scalpel and tweezers (or punch tool)''' | ||
| - | *''' | + | *'''ddH20''' |
*'''99% ethanol''' | *'''99% ethanol''' | ||
*'''0.5 ml tubes''' | *'''0.5 ml tubes''' | ||
<br> | <br> | ||
*Put 10 µl of pre-warmed TE into a 0.5 ml tube. | *Put 10 µl of pre-warmed TE into a 0.5 ml tube. | ||
| - | *Cut paper spots using scalpel and tweezers (or punch tool, following | + | *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. | *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. | *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. | ||
Latest revision as of 12:52, 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
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.
