Red Blood Cell (Hypotonic) Lysis Buffer

Simon Mauch Simon.Mauch at uni-konstanz.de
Wed Jun 24 15:16:23 EST 1998


Hi Bert, 
My protocol for lysis of red blood cells:
Pellet the the blood cells and resuspend in a Tris-NH4Cl solution (0.1
ml packed cells/ml Tris-NH4Cl). Hold at room temperature for 5 to 10
min.
Centrifuge at 300xg for 10 min. (You might underlay the cell
suspension with FCS before centrifugation)
Repeat the process if red blood cells are evident in the pellet.
Wash the pellet with PBS.
Use the Pellet for DNA isolation

stock solutions:
0,16 M NH4Cl (8.3 g/l)
0,17 M Tris-HCl, pH 7.6
working solution:  9 parts NH4Cl stock solution and 1 part
Tris-solution.  Adjust the pH to 7,2 with HCl

Simon




>Colleagues,
>
>For a DNA extraction at work, it would be good to be able to mix up
>a hypotonic buffer capable of lysing red cells, but keeping white 
>cells (or at least nuclei) intact.  Anybody know a recipie for
>such a salt-water mix?
>
>Bert Gold
>Sherman Oaks, California
>http://www.ktb.net/~bgold




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