RBC prep

Michael Gerdes mg690769 at bcm.tmc.edu
Fri Aug 11 08:52:54 EST 1995


In article <kfischer-1008950332210001 at duckter.glaxo.med.ualberta.ca> kfischer at gpu.srv.ualberta.ca (Karl Fischer) writes:
>Hmm...if you have a variable speed microfuge you might want to experiment
>with Ficoll-Hypaque; PBMC's should band while RBC pellet out.


I used to do a lot of RBC preps for membrane isolation.
I used a cooled clinical C-fuge, but there is no reason why a microfuge 
won't work, provided that you can set the speed way down ~3000rpm.

Spin for 15-20min @4C,
remove plasma and white buffy coat (this is where the white cells are)
wash in isotonic buffer, prechilled to 4C
respin.
repeat several times.

While I don't know what sort of volume you will need to MT DNA isolation, 
this should work.  Try several speed settings before doing a large scale 
prep.

The trick is to avoid lysis of the RBC's.  Fresh blood is important.

Hope this helps-

MiJoGe
mg690769 at mbcr.bcm.tmc.edu
Baylor College of Medicine






More information about the Methods mailing list