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