1. Add 20 ml of sterile PBS to the blood smear and scrape 1/3 of slide with a razor blade.

2. Add to a microcentrifuge tube, wash the area with an additional 30 ml and combine.

3. Vortex and let stand overnight at room temperature.

4. Boil for 10 minutes to complete cell lysis.

5. Centrifuge at maximum speed in a microfuge for 1 minute.

6. Transfer supernatant to fresh freezing vial for storage. Store at -80°C.

7. Use ≤ 10 ml for 50 ml PCR reactions. Dilute 1:2-1:10 if the smear is thick and the PCR reaction indicates the presence of inhibitors.