Electroelution of agarose fragments Electroelution buffer 1 M Tris, pH 7.5 12.0 mls 0.5 M EDTA 0.24 mls 1 M NaCl 3.0 mls qs to 600 mls dH2O Acetate cushion 3 M NaAcetate pH 4.8 480 ul 0.1 % Bromphenol Blue 40 ul 1. Place gel slices in trough 2. Remove all air bubbles, then layer 80 ul of acetate cushion 3. Electroelute at: 120V for ~1Kb to 140V for >2.5Kb for 40 min for ~1Kb to 60 min for >2.5Kb 4. Collect ~300 ul of salt cushion, add 3X volumes of 95% ethanol to precipitate 5. Remove gel slices Clean wells Run for 10 min longer Clean wells again Rinse thoroughtly to remove any extraneous DNA