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        Phenol/chloroform Extraction of DNA

        互联网

        1199

        Materials:

        phenol:chloroform (1:1)

        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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