小中大
这是 Nycoprep A 的 protocol
1. collect blood into a tube containing anticoagulant(EDTA,heparin,ACD)
2.Dilute the blood by addtion of an equal volume of 0.9%Nacl.
3. Carefully layer 6mL of the diluted boole over 3 mL Nycoprep A in a 12-15mL centrifuge tube.Avoid mixing of bllod and tube to prevent the formation of aerosols.
4.Centrifuge at 600×g for 20 min.
5.The mononuclaer cells can be harvested from the interface between the plasma layer and the Nycoprep A solution using a Pasteur pipette.
6.Transfer the cells to a smaller tube ,and add some Tris-BSS or a similar physiological solution,and Centrifuge at 400×g for 10 min
7. Resuspend the pellet and spin down again.
