Prepare DNA Extraction Buffer (DEB):
0.1M Tris-HCl (pH 8) 4.5 mL of 1.0 M
0.1M Na-EDTA (pH 8) 9 mL of 0.5M
0.1M KH2PO4 (pH 8) 0.54 g
0.8M Guanidine HCl 3.44 g
0.5% Triton-X 100 0.225 mL (225 μL) of 100%
Add above ingredients to 50 mL tube.
Add milli-Q water to ~40 mL
Add NaOH to pH 10 (several drops at a time)
Add milli-Q water to 45 mL
Filter-sterilize to remove possible spores
Autoclave. Slightly loosen lid so that it is not air-tight. Recover from autoclave very soon after the autoclave cycle is completed.
Pour autoclaved solution into fresh 50 mL tube.
Aliquot into 1.5 mL tubes.