RNA REMOVAL: Help!
ez001427 at dale.ucdavis.edu
ez001427 at dale.ucdavis.edu
Thu Aug 25 18:50:08 EST 1994
Dear Netters,
I could use some help in removing trace quantities of RNA from a
pBluescript plasmid prep that I have. I made a very large (15 mg) prep
by traditional means, and used a combination of RNASE A and RNASE T1 to
chew up the RNA. I thought that that would be sufficient, but it turns
out that the small amount of tiny (runs below 200 bp vs. dsDNA ladder)
RNA remaining is a problem. So I used a CsCl2 gradient, which removed
90 % the RNA, but still there is enough to cause a problem. I would
appreciate any suggestions for chemically gentle (non-DNA-damaging)
methods to remove the trace RNA contamination that remains.
One suggestion I have had is to run a gel filtration column. If
any of you have experience with such, and could recommend the best media,
column conditions, etc.. I would be very grateful.
Thank you in advance for your help, either as a followup post
here or as email to the address below.
Marc Goldstein
Section of Plant Biology
U. C. Davis
magoldstein at ucdavis.edu
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