Prehybridization Treatment and Hybridization

1. Proteinase K is prepared as a 10 mg/mL stock and is stored at -20°C in small aliquots. On day of use dilute to 10 pg/mL with PBS.

3. 4% Paraformaldehyde prepared as in Subheading 2.2.

4. Methanol.

5. Hybridization solution: 50% deionised formamide (BDH/Merk Ltd, Poole, UK-molecular biology grade); 5X SSC (pH 4.5 with citric acid [BDH]); 50 pg/mL yeast tRNA; 1% SDS; 50 pg/mL heparin (Sigma). Store at room temperature for up to 1 mo.

6. Histolene.

7. 20X SSC: 20-fold stock solution of 0.15 M NaCl and 0.015 M sodium citrate.

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