DNA isolation from frozen blood

Jerry Kropp jkropp at itsa.ucsf.edu
Thu Oct 17 20:50:14 EST 1996


In article <3263DE48.4EC8 at med.kuleuven.ac.be>, Peggy De Clercq
<peggy.declercq at med.kuleuven.ac.be> wrote:

> Hello,
> 
> I have to isolate DNA from a lot of 1 mL frozen blood samples. I need 
> that DNA for PCR. Does there exist a rapid method to isolate DNA from 
> only 1 mL blood?
> 
> Thanks
> Peggy De Clercq

When whole blood is frozen the RBCs lyse, but the white cells don't. Of
course the DNA is in the whites. Spin the thawed material at 5000g for 5
min and carefully remove an discard the sup. It's hard to see the WBC
pellet at this point because of the hemolysate, so don't try to get all
the sup at first. Wash twice with normal saline and the pellet will be
easily visible.
You can use any standard method for isolating DNA from cultured cells
after that. I still use the old-fashioned phenol-chloroform, but I'm a
Luddite.
Good Luck,
Jerry



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