DNA Isolation from Lymphocytes

1. 1X Blood lysis buffer (1 L): 155 mM NH4Cl , 10 mM KHCO3, 1 mM EDTA, pH 7.4. This solution should be stored at 4°C up to 6 mo. Blood should be handled using universal precautions. Take care to discard all blood contaminated materials in appropriate biological waste containers.

2. 1X Nucleus lysis buffer (1L): 10 mM Tris-HCl, 400 mM NaCl, 2 mM EDTA, pH 8.2. Store at 4°C up to 1 y.

3. 10% SDS Ultra PURE (Gibco BRL). Store at room temperature up to 1 y.

4. Pronase 20 mg/mL (Sigma). Store at -20°C up to 1 y.

5.6 M NaCl. Store at room temperature up to 6 mo.

6. Absolute ethanol.

7. TE-4 (0.5L): 10 mM Tris-HCl, 0.1 mM EDTA, pH 7.5. Store at room temperature up to 1 y.

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