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Template:Team:CityColSanFrancisco/Notebook/Notebook/CCSF Protocols
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BASIC PROTOCOL FOR ACCESSING iGEM PARTS DISTRIBUTION DNA<br> | BASIC PROTOCOL FOR ACCESSING iGEM PARTS DISTRIBUTION DNA<br> | ||
1. Find desired part at partsregistry.org <br> | 1. Find desired part at partsregistry.org <br> | ||
| - | 2. Click "get this part" to find distribution plate number and coordinates. | + | 2. Click "get this part" to find distribution plate number and coordinates.<br> |
| - | 3. Clean foil surface of desired plate with a little ethanol. | + | 3. Clean foil surface of desired plate with a little ethanol. <br> |
| - | 4. Pierce through foil surface with sterile pipet tip (on a pipet of course) containing 15uL sterile dH2O. | + | 4. Pierce through foil surface with sterile pipet tip (on a pipet of course) containing 15uL sterile dH2O.<br> |
| - | 5. Pipet up & down a couple of times to dissolve the DNA and buffer in the dH2O. | + | 5. Pipet up & down a couple of times to dissolve the DNA and buffer in the dH2O.<br> |
| - | 6. Pipet the plasmid dilution into a labelled, sterile 1.5mL tube | + | 6. Pipet the plasmid dilution into a labelled, sterile 1.5mL tube.<br> |
| - | 7. Put the labelled tube into the CCSF DNA box, and note the box coordinates into which the tube is placed. | + | 7. Put the labelled tube into the CCSF DNA box, and note the box coordinates into which the tube is placed.<br> |
| - | 8. Document the location of the tube on the Google doc "CCSF DNA box index", found at http://spreadsheets.google.com/ccc?key=t0CX5NWkXQL0_8v-w2MZaGA&inv=dirkvandepol%40gmail.com&t=7890798853979134080 | + | 8. Document the location of the tube on the Google doc "CCSF DNA box index", found at http://spreadsheets.google.com/ccc?key=t0CX5NWkXQL0_8v-w2MZaGA&inv=dirkvandepol%40gmail.com&t=7890798853979134080<br> |
Latest revision as of 19:42, 9 July 2009
BASIC TRANSFORMATION PROTOCOL FOR USING CCSF COMPETENT CELLS
1. Thaw competent cells on ice for 10 min.
2. Pipet 200uL cells into chilled 1.5mL rube containing 1 uL plasmid of interest
3. Mix gently by tapping
4. Incubate cells on ice in presence of plasmid for ~20 min.
5. Heat shock at 42C for 90 sec.
6. Return to ice for 2 min.
7. Plate 100uL of transformation onto LB-amp plate
BASIC PROTOCOL FOR ACCESSING iGEM PARTS DISTRIBUTION DNA
1. Find desired part at partsregistry.org
2. Click "get this part" to find distribution plate number and coordinates.
3. Clean foil surface of desired plate with a little ethanol.
4. Pierce through foil surface with sterile pipet tip (on a pipet of course) containing 15uL sterile dH2O.
5. Pipet up & down a couple of times to dissolve the DNA and buffer in the dH2O.
6. Pipet the plasmid dilution into a labelled, sterile 1.5mL tube.
7. Put the labelled tube into the CCSF DNA box, and note the box coordinates into which the tube is placed.
8. Document the location of the tube on the Google doc "CCSF DNA box index", found at http://spreadsheets.google.com/ccc?key=t0CX5NWkXQL0_8v-w2MZaGA&inv=dirkvandepol%40gmail.com&t=7890798853979134080
