DNA from Whole Blood

Jerry Kropp jkropp at itsa.ucsf.edu
Mon Nov 18 17:16:42 EST 1996


In article <56pn3n$dj0_001 at leeds.ac.uk>, MSJSAR at leeds.ac.uk (Steve Rose) wrote:

> I have been given frozen samples of blood which have already had their 
> lymphocytes removed by density gradient centrifugation. What's left is mainly 
> Red Blood cells and Polymorphs. Obviously RBC's dont have any DNA, but the 
> POlymorphs do. Does any one have a method for extraction DNA clean enough for 
> PCR from these frozen samples?
> 
> Thanks for any help.
> 
> Steve.
> 
Hi Steve'
Freezing does not usually lyse the WBCs-only the RBCs. Just thaw and spin
@~5000g and carefully aspirate the dark hemolysate so it doesn't mask the
WBC pellet. Wash the lysate out with normal saline and the pellet will
become east to see. Then use any std. DNA extraction protocol. I still use
good ol' phenol/proteinase K.
Good Luck, 
Jerry



More information about the Methods mailing list