| 1. | Place volume of cells (nucleated) obtained from ficoll enrichment equal to ~108 cells into amplification tube. |
| 2 | Wash cells 1X with normal saline. |
| 3. | Discard the supernatant following gentle (ie <1000 X g) centrifugation. |
| 4. | Resuspend cell pellet with 20ul of GeneReleaser®. |
| 5. | Vortex |
| 6 | Overlay with ~50ul mineral oil. |
| 7. | Lyse cells + GeneReleaser® with either thermocycle or microwave procedure in amplification tubes. |
| 8. | Add 100ul of sterile DI to the lysate. |
| 9. | Vortex. |
| 10 | Centrifuge 10,000 X g for 5 minutes. |
| 11. | Transfer supernatant to fresh tube. |
| 12. | Use 1-10ul as source of target DNA for amplification. |
| 13. | Supernatant recovered in step 11 can be stored for at least 1 year at -20°C. |
