Quick Preparation of Plasmid DNA from Yeast

Adapted from A. Lorincz (BRL)

Last update 6/29/99

 

  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

 

Fred Hutchinson Cancer Research Center
1100 Fairview Ave. N. PO Box 19024 Seattle, WA 98109
©2014 Fred Hutchinson Cancer Research Center, a 501(c)(3) nonprofit organization.
Terms of Use & Privacy Policy.