Materials see Note 4 21 Dephosphorylation

1. DNA: either a purified DNA fragment in water, IX TE, or a restriction digest mixture after incubation with the appropriate restriction enzyme (see Chapter 50).

2. 2X CIP buffer: 2 mMZnCl2, 2 mMMgCl2, and 20 mMTris-HCl, pH 8.3

3. Calf intestinal alkaline phosphatase (or bacterial alkaline phosphatase). CIP and BAP are often supplied as ammonium suspensions. The ammonium ions must be removed by dialysis with IX TE before use.

4. Phenol saturated in IX TE.

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