Buccal cell DNA preps

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 ul 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 95C 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 ul 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 ul of supernatant for standard PCR.

From Chromaffin Cell & Hypertension Research Department at UCSD