Template:Team:CityColSanFrancisco/Notebook/Notebook/CCSF Protocols
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Dirkvandepol (Talk | contribs) (New page: <Br> 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 interes...) |
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- | BASIC TRANSFORMATION PROTOCOL FOR USING CCSF COMPETENT CELLS | + | BASIC TRANSFORMATION PROTOCOL FOR USING CCSF COMPETENT CELLS<br> |
- | 1. Thaw competent cells on ice for 10 min. | + | 1. Thaw competent cells on ice for 10 min.<br> |
- | 2. Pipet 200uL cells into chilled 1.5mL rube containing 1 uL plasmid of interest | + | 2. Pipet 200uL cells into chilled 1.5mL rube containing 1 uL plasmid of interest <br> |
- | 3. Mix gently by tapping | + | 3. Mix gently by tapping <br> |
- | 4. Incubate cells on ice in presence of plasmid for ~20 min. | + | 4. Incubate cells on ice in presence of plasmid for ~20 min. <br> |
- | 5. Heat shock at 42C for 90 sec. | + | 5. Heat shock at 42C for 90 sec. <br> |
- | 6. Return to ice for 2 min. | + | 6. Return to ice for 2 min. <br> |
- | 7. Plate 100uL of transformation onto LB-amp plate | + | 7. Plate 100uL of transformation onto LB-amp plate <br> |
Revision as of 19:23, 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