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        Phenol/chloroform Extraction of DNA

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        777

         

        Materials:

        bullet phenol:chloroform (1:1)
        bullet chloroform
        1. Add an equal volume of buffer-saturated phenol:chloroform (1:1) to the DNA solution.
        2. Mix well. Most DNA solutions can be vortexed for 10 sec except for high molecular weight DNA which should be gently rocked. (If using Phase-Lock Gel, follow procedure M.1 )
        3. Spin in a microfuge for 3 min.
        4. Carefully remove the aqueous layer to a new tube, being careful to avoid the interface.
        5. (Steps 1-4 can be repeated until an interface is no longer visible)
        6. To remove traces of phenol, add an equal volume of chloroform to the aqueous layer.
        7. Spin in a microfuge for 3 min.
        8. Remove aqueous layer to new tube.
        9. Ethanol precipitate the DNA

         

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