ALPF Medical Research

DNA Extraction from Fin Biopsies for PCR or Southern Analysis

1. Place fin biopsy in 500 pL of 10 mM Tris-HCl, pH 8.0, 10 mM EDTA, 100 mM NaCl, 0.5% SDS, and 200 pg/mL proteinase K at 55°C for 12 h.

2. Add an equal volume of phenol:chloroform (1:1). Mix gently.

3. Spin in microfuge for 5 min.

4. Remove the upper, aqueous layer to a fresh microfuge tube.

5. Repeat steps 2-4.

6. Extract with an equal volume of chloroform.

7. Add 1/10 vol of 2.5 M sodium acetate, pH 5.5, and 2.5 vol of ethanol.

8. Spin down in microfuge for 10 min.

9. Rinse pellet in 80% ethanol and respin.

10. Dry the pellet.

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