Genomic DNA isolation from mammalian cells

Deitiker, Philip R via methods%40net.bio.net (by pdeitik from bcm.tmc.edu)
Wed Apr 22 12:47:52 EST 2009


>From Whole Blood, Qiagen Gentra PureGene DNA Isolation Kit. This will get Genomic DNA. 
One precaution on its use, fresh RBCs are hard to lyze, it requires washing in RBC lysis buffer and carefully drawing off the RBC layer from the White blood cells. Small levels of RBC contamination at the point of DNA extraction (White blood cell lysis and solubilization) can result in Fe/Heme that
can really screw up the DNA prep. Blood should be collected in Acid Citrate Dextrose(ACD, BD Yellow Top Tubes) and 8 mls is more than enough for most efforts. For blood the kit is reliable.  
 



-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu [mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of Dwayne Taliaferro
Sent: Tuesday, April 21, 2009 9:51 PM
To: methods from magpie.bio.indiana.edu
Subject: Genomic DNA isolation from mammalian cells

Does anyone have a quick and reliable protocol for isolating gDNA from
mammalian cells?  I  would be interested.

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