This is a simple gel extraction protocol that does not require any expensive spin kits. If you want to see a protocol for the Qiagen QiaQuick spin kits see Gel extraction. Also for dialysis gel extraction, see Gel extraction again for a detailed protocol.
Fragment Purification: Agarose
1. Run DNA on "Low melt" agarose gel: 1% -600 bp to 5kb; 2% 300700bp; Smaller? Use 2% "Nusieve" + 1% "Low melt".
2. After band have separated, visualize band on UV box (minimize exposure of DNA to UV), cut band out (DO NOT scratch filter on box).
3. Add 100l of T.E. - Tris-EDTA (10mM Tris-HCl pH 7.6, 1mM EDTA) to band, crush, heat to 65oC for approx. 5 min, add 200l of phenol, vortex, heat 65oC 3 min., vortex
4. Microfuge 5 mins, remove supernant
5. Add 100 l of T.E. to phenol, vortex, heat 65oC 3 min. vortex
6. Microfuge, pool supernants.
7. Chloroform extract (approx. 400 ul), microfuge 3 min.