| 1. | Transfer an aliquot of whole blood containing a population of 106 nucleated cells (~200µl) to a 0.5 µl amplification tube. |
| 2. | To the aliquot of whole blood add an equal volume of 1% Triton X-100 (1% wt/volume prepared in sterile H2O). Vortex 1-2 seconds and then place on ice or at 4°C for 5 minutes. |
| 3. | Remove the tubes form 4°C and microfuge for 1 minute at 12,000 xg. |
| 4. | Remove and discard the supernatant containing the RBC lysate. |
| 5. | Wash the remaining cell pellet 2 x with 200 µl of 1x amplification buffer by resuspending the cells in the buffer and repeating the centrifugation step. |
| 6. | Following the last wash step, discard the supernatant as completely as possible. |
| 7. | Add 20µl of GeneReleaser® and proceed with either the thermocycle or microwave lysis protocols. |
