1. Wash cells (105-106) with PBS, harvest, and transfer to 1.5 ml
microcentrifuge tube.
2. Resuspend cell pellet in 0.5 ml of the lysis buffer (5 mM Tris-HCl pH 8.0,
20 mM EDTA, 0.5% Triton X-100).
3. Incubate on ice for 20 minutes.
4. Centrifuge at maximum speed for 30 minutes.
5. Transfer the supernatant to a fresh tube.
6. Extract with 0.5 ml of phenol:chlorofrom:isoamyl alchohol (25:24:1).
7. Add 1/10 volume of 3M sodium acetate and precipitate with 2 volumes of
ethanol.
8. Centrifuge to pellet DNA.
9. Wash the pellet with 70% ethanol and air dry.
10. Resuspend the DNA in 30-50 ml of
TE/10 mg/ml RNase A.
11. Detect the DNA ladder by running on a 1.2% agarose gel.
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