Quick Preparation of Plasmid DNA from Yeast

 

 

 

1. Pick a medium size yeast colony (~3 mm dia) and transfer to 200 ul of lysis buffer.
2. Add an equal volume of glass beads (0.45 mm dia). Mix on vortex at top speed for 1 min.
3. Add 200 ul of phenol/CHCl3 (2/1). Extract one time.
4. Ethanol precipitate DNA. Wash once with 80% ETOH.
5. 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