Can you please write about DNA extraction from blood andpreparation of blood samples for DNA extraction. This is theprotocol:
1.Take 750 μl of blood from each blood sample, and transfer itto new 1.5ml tube;
2. Mix 750 μl of blood with 1ml RBC lysis buffer, respectively(avoid stock contamination by not touching the walls of tube bypipette tip);
3. Centrifuge at 2300 rpm for 5 minutes;
4. Take out the liquid, keep the pellet;
5. Mix 1 ml of RBC lysis buffer with pellet in 15ml centrifugetube;
6. Add PBS up to 14 ml by pipette controller (increase volume andcontrast);
7. Centrifuge at 2200 rpm, at 4°C, for 5 minutes
8. Discard the supernatant, keep the pellet;
9. Add 1 ml of RBC lysis buffer;
10. Add PBS up to 10 ml;
11. Centrifuge at 2200 rpm, at 4°C, for 5 minutes;
12. Discard the supernatant and resuspend the pellet in 1 ml ofTrizol;
13. Store the samples at -20°C
Can you explain the protocol, why are we doing these steps. Alsoif you can say the function of each chemical:
- RBC lysis buffer
- PBS
- Trizol
Thank you in advance. I really really need your help.
