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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