Quick Preparation of Plasmid DNA from Yeast
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- Pick a medium size yeast colony (~3 mm dia) and transfer to 200 ul of lysis buffer.
- Add an equal volume of glass beads (0.45 mm dia). Mix on vortex at top speed for 1 min.
- Add 200 ul of phenol/CHCl3 (2/1). Extract one time.
- Ethanol precipitate DNA. Wash once with 80% ETOH.
- Dry DNA and resuspend in 100 ul TE and use to transform E. coli (try 1.0 and 0.1 ul DNA).
Lysis Buffer (1 ml)
| 100 mM NaCl | 100 ul 1 M NaCl |
| 10 mM Tris 8.0 | 10 ul 1 M Tris 8.0 |
| 1 mM EDTA | 4 ul 0.25 M EDTA |
| 0.1% SDS | 10 ul 10% SDS |
| 876 ul H2O |
