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. Author manuscript; available in PMC: 2015 May 5.
Published in final edited form as: Methods. 2004 Jul;33(3):199–205. doi: 10.1016/j.ymeth.2003.11.014

Table 4.

Smash and Grab rescue of plasmids from yeast to E. coli

1. Allow colonies to form on the S. pombe transformation plate (4–5 days after plating). Collect the cells by gentle scraping of the plate in 1 ml sterile water using a glass spreader. Transfer cells to a microfuge tube and concentrate by a brief centrifugation.
2. Resuspend the cell pellet in an equal volume of Smash and Grab buffer (1% SDS; 2% Triton X-100; 100 mM NaCl; 10mM Tris, pH 8.0; and 1 mM EDTA). Transfer 200 μl of cells to a new tube and add 200 μl phenol–chloroform and 300 μl acid washed glass beads. Vortex for 5 min to lyse >50% of the cells as determined by microscopy.
3. Pellet cell debris by microfugation for 5 min. Remove 50 μl of the top (aqueous) layer to a new microfuge tube. Avoid material close to the interface that may be contaminated with phenol–chloroform.
4. Add 50 μl isopropanol, precipitate on ice for 10 min, and pellet DNA by microfugation for 10 min. Remove the liquid and allow pellet to air-dry. Resuspend in 5 μl sterile water.
5. Transform electroporation-competent E. coli strain XL1-Blue (Stratagene) with 1 μl DNA according to manufacturer's instructions (in a slight modification we use 10 μl competent cells in 89 μl sterile water for electroporation and plate the entire transformation onto a single LB Amp plate).