The preparation of Genomic DNA from Thermus aquaticus was performed using a modification of the method proposed by Hoffman and Winston (1987) for the preparation of yeast DNA. This technique is outlined below:
Grow 10 ml of culture to saturation in the appropriate media.
Collect the cells by centrifugation. Remove the supernatant.
Add 200 ul of TE and vortex until cells are in solution. Transfer to 1.5 ml microfuge tube.
Add 200 ul of lysis buffer (2% Triton X-100, 1% SDS, 100 mM NaCl, 10 mM Tris-Cl (pH 8), 1 mM EDTA).
Add 200 ul phenol:chloroform.
Add 0.3 g of acid-washed glass beads.
Vortex 3-5 minutes.
Microfuge 10 minutes and collect the aqueous layer which contains the DNA.
Subsequent purification by ethanol precipitation is strongly recommended.
Reference:
Hoffman and Winston, (1987) Gene 57:267-272.