Important: Extract the DNA within one week of receiving samples. Samples that have been processed should be frozen to prevent degradation. Freeze samples between use each day.
1. Add 600 ml of 50 mM NaOH to a 1.5 ml eppendorf tube and insert brush in tube. (Clip off handle of brush with wire cutters so tube can be closed. Cutters should be rinsed with EtOH between samples to prevent contamination.)
2. Vortex thoroughly to mix, for at least 10 s.
3. Heat to 95∫C for 5 min.
4. Spin briefly to pool condensation.
5. Remove brush with tweezers (tweezers should be rinsed with ethanol between samples to prevent contamination) and add 60 ml 1M Tris pH 8.0 to tube. Vortex to mix for at least 10 s.
6. Spin for 1 minute at 13,000 rpm and decant supernatant to a clean storage tube.
7. Use 2-5 ml of supernatant for standard PCR.
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