Materials

1. Sylgard (Dow Corning, Barry, South Glamorgan, UK): This is made up 9 parts to 1 with a hardener. Sylgard dissolves in solvents, such as Histoclear and xylene.

2. Agarose-filled dishes: Use 1% agarose in distilled water. Heat to >60oC to dissolve and cool to set.

3. Cleaned coverslips: Wash in 70% ethanol/1% conc. HCl and then 2% (TESPA) 3-aminopropyl- triethoxysilane/3-(triethoxysilyde)-propylamine for 10 min. Rinse in acetone for 10 min and then distilled water before drying.

4. Gelvatol (Monsanto Chemicals, Knowsley, UK)/ Mowiol (Hoechst, AG, Frankfurt, Germany): Use 2.4 g Mowiol to 6 g of glycerol. Stir to mix. Add 6 mL water and leave for several h at room temperature. Add 12 mL of 0.2 M Tris buffer (pH 8.5), and heat to 50oC for 10 min to dissolve. For fluorescence, add 1,4-diazobicyclo-(2.2.2)-octane (DABCO).

Was this article helpful?

0 0

Post a comment