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Quick Yeast DNA Prep: Isolation of Total DNA (genomic and plasmid)

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Quick Yeast DNA Prep: Isolation of Total DNA (genomic and plasmid)

Linda Hoskins, Hahn Lab 10/16/98

  1. Grow a 5 ml YPD O/N culture inoculated with a single yeast colony at 30 deg.
  2. Transfer culture to a small 13 x 100 glass tube. Spin down cells 2 min. in tabletop centrifuge. Pour off supernatant.
  3. Wash cells with 5 ml H2 O. Spin down cells 2 min. Pour off supernatant. Cell pellet can be stored at -20 deg.
  4. Resuspend cells in 500 ul lysis buffer by vortexing.

     

    Lysis buffer:   20 ml:
    0.1M Tris pH 8.0   2 ml 1M Tris 8.0
    50 mM EDTA   2 ml 0.5M EDTA
    1% SDS   2 ml 10% SDS
        14 ml H2 O
         

 

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