Genomic DNA

Tracy G. Stevens tstevens at shrinkatron.utmb.edu
Wed Feb 22 15:35:19 EST 1995


I am having trouble getting genomic DNA out of HEMS cells.  I have had no 
trouble extracting DNA from HL-60's, PA317's, NIH 3T3's, & PE501's.  I am 
using the same method (Round Method - lysis buffer is SDS, EDTA, Tris, and 
RNase for 2 hrs at 37, ProtK for 1 hr at 37, room temp overnight, followed by 
phenol-chloroform extractions, 1/10th volume Na Acetate topped by 2 volumes 
EtOH, and spooling out the DNA.  Not technically demanding), fresh reagents, 
old reagents, more gentle mixing to no avail.  My best yield was 400ug from 5 
of the T-150 flasks.  In other words, not much.

Does anyone have any suggestions, words of wisdom or encouragement?

Thanx in advance for any and all replies.



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