re "in vitro transcr...."

S. S.
Thu May 18 03:39:31 EST 1995


In article <3pdkrs$jip at news.duke.edu> Meyer Lab writes:
> We tried to inject poly A-tailed mRNA (invitro transcribed) into
>eukariotic cells. However the cells died. We assume that there may be some
>salt left from the poly A tailing step (mainly Mn). Does anybody have
>experiences with in vitro transcribed mRNA in euk cells and does somebody
>know a RNase free system (most commercial avaiable columns are not RNase
>free) to get rid of theses ions?
>Thanks Thomy

I'd try to dialyze the samples.  For microscale I use circullar
~25mm filters from Millipore, Catalogue No. VSWP 025 00 (no 
connections with the company).  Just put the filter on the surfice
of water or desired buffer, place aliquots (5-50ul) and collect 
them after few minutes.  Sorry, but I don't know whether they are
free of RNAses.  Cheers.  Slawek.



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