Protocol: DNA extraction from blood samples 1. Transfer 80l of blood sample to a new eppendorf. 2. Add 1ml of NH4Cl to each tube, mix vigorously by inverting the tube and incubate at room temperature for 10 minutes. 3. Centrifuge for 5 minutes at 2000rpm at 4ºC. 4. Remove the supernatant. 5. Add 1ml of PBS and mix. 6. Centrifuge for 5 minutes at 2000rpm at 4ºC. 7. Remove PBS. 8. Ressuspend the pellet in the remaing PBS (50l). 9. Make a solution mix of lysis buffer (3ml) and proteinase K (3l). 10. Add 200l of the solution mix to each tube. 11. Incubate at 50ºC with mix (300rpm) during many hours. 12. Add 200l of Phenol:Chloroform Isoamyl Alcohol to each tube and mix vigorously by inverting the tube. 13. Centrifuge at maximum speed (13200) for 3 minutes. 14. Transfer the aqueous fase (top) to a new tube. 15. Add 200l of Isopropanol and mix by inverting the tube. 16. Centrifuge at 13200 rpm for 20 minutes. 17. Remove the supernatant. 18. Add 1ml of alcohol 70%. 19. Centrifuge at 13200 rpm for 10 minutes. 20. Remove the supernatant. 21. Dry the pellet at room temperature. 22. Add 20l of TE buffer to each tube. 23. Incubate at room temperature for 4h. 24. Determine concentration.
Pages to are hidden for
"Protocol DNA extraction from blood samples (DOC)"Please download to view full document