1- Stain gel with Ethidium bromide solution.
2- Wash gel in 2x distilled water.
3- Take picture.
4- Soak gel 2 x 15’ in ~ 250 ml 0.25 N HC1
5- Rinse in water
6- Soak gel 2 x 15’ in 0.5N NaOH – 1 M NaCl solution.
7- Rinse in water.
8- Soak for 30’ in 0.5 M Tris – 1.5M NaCl solution.
9- Transfer the DNA to a nylon membrane using 10 x SSC or 20 x SSC. The nylon membrane should be UV-irradiated afer blotting.
10- If nitrocellulose membrane is used, soak in 2X SSC before transfer.
SOLUTIONS
0.5 N NaOH (26.16 ml conc NaOH/liter).
1.0 M NaCl (58.45 g /liter).
0.25N HC1 (20 ml conc. HCl/liter).
0.5 M Tris base (60.57 g /liter)
1.5 M NaCl (87.67 g / liter)