| Contributor: |
The Laboratory of Jasper Rine at the University of California, Berkeley
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| This protocol is a streamlined version of the lithium acetate/PEG transformation protocol used to introduce DNA into yeast. |
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1. Start with 0.5 ml of a yeast culture (any phase) or 1 mm3 of cells taken from a plate with a sterile toothpick.
2. Add the sample of plated yeast to 0.5 ml of ddH2O in a microcentrifuge tube and mix until the cells are in suspension.
3. Pellet the cells with a brief centrifugation in a microcentrifuge at full speed and remove the supernatant.
4. Add 1 to 5 μg of the transformation DNA and 15 μl of Salmon Sperm DNA Carrier Solution and mix.
5. Add 0.5 ml of Plate Solution.
6. Incubate the mixture at room temperature for about 14 hr.
7. With a yellow pipette tip, pull out the bottom 50 μl from the tube and plate out these cells.
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| Salmon Sperm DNA Carrier Solution |
| 7 mg/ml Sonicated Salmon Sperm DNA
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| Plate Solution |
| Autoclave and cool to room temperature
40% (w/v) PEG-3350
1 mM EDTA
10 mM Tris, pH 7.5
0.1 M Lithium Acetate
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Polyethylene Glycol (PEG) 3,350
Tris
Lithium Acetate
EDTA
DNA, Salmon Sperm
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No hints are associated with this bioProtocol
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