Re-use of magnetic beads in Promega's PolyA selection system
Lely Chow
lelyc at u.washington.edu
Mon Oct 30 20:18:27 EST 1995
Hi Everyone!
Would someone like to comment on the re-use of the streptavidin-coated
magnetic beads for polyA selection? I did this by mistake, and I found
out that I now risk 1) reduced yield of polyA product; and 2)
contamination of my mRNA with previous mRNA. It seems as though my
yields were not detectably reduced, which makes sense since I did not
saturate the capacity
of the beads the first time. It is also possible that what little RNA was
left over from a previous selection may not contribute greatly to the new
batch of RNA, or that it has degraded with time (25 days at 4 oC). I
add approx. 60 ug of RNA to each tube (max.that can be added is 1 mg), and I
reused
each tube twice. I am using the RNA on polyA dot blots to compare three
treatment conditions.
It would be nice to know whether I need to worry about contamination, since
it is very difficult for me to
get the RNA I need for these experiments, and I would like to avoid
having to redo the experiments.
Hope someone has some helpful comments on this matter. Thanks for your time.
--Lely Chow
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