Preparation of Template for Riboprobe Synthesis

1. Appropriate restriction endonucleases and manufacturer's buffers (e.g., Boehringer Mannheim).

2. Agarose (Life Technologies).

3. 10X TBE: 0.89 M Tris-borate, 0.89 M boric acid, 0.02 M EDTA; per liter 108 g Tris base, 55 g boric acid, 9.4 g EDTA.

4. Ethidium bromide (10 mg/mL). Stored at 4°C in the dark. Caution: ethidium bromide is a potent carcinogen.

5. Phenol (molecular biology-grade; Life Technologies). Melted at 65°C in a fume cupboard and equilibrated with 0.1 M Tris-HCl, pH 7.0 followed by addition of a few crystals of 8-hydroxyquinoline, which will color the solution yellow. Stored in the dark at 4°C and discarded if the solution begins to turn orange.

6. Chloroform (AnalaR; Merck BDH).

8. Absolute (100% v/v) ethanol and 70% (v/v) ethanol.

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