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        Yeast DNA Prep

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        Protocol

        1. grow up yeast culture to appropriate density (near saturation)
        2. spin 1.5 mls of culture for 1 min in microfuge and aspirate off supernatant
        3. resuspend pellet in 200 ul breaking buffer
        4. wear gloves and add:
          • 200 ul phenol:choloroform:isoamyl alcohol (25:24:1)
          • 200 ul (@200 mg) glass beads
        5. close cap tightly and vortex for 2.5 min.
          • Be careful when vortexing; label can be dissolved by the phenol.
          • Hold cap tightly so it doesn''t open or spill.
        6. add 200 ul TE buffer and spin for 5 min, in microfuge
        7. transfer 350 ml aqueous (top) layer to fresh eppendorf.
        8. add 1 ml 95% ethanol and mix well, let sit for 10 minutes
        9. spin for 2 min, take off supernatant, and let dry upside down 10 min.
        10. resuspend pellet in 50 ul TE buffer or water.

        You can now use 1-2 ul of this crude yeast plasmid DNA prep to transform E. coli.

        Materials

         

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