Phosphorothioate oligo questions
g086456 at JAGUAR1.USOUTHAL.EDU
Fri Jan 20 18:11:33 EST 1995
On Fri, 20 Jan 1995, Karen Gravitt wrote:
> Hello everyone,
> I have a couple of questions reguarding anti-sense phosphorothioated
> oligonucleotides. I will be using 20-mers in tissue culture. First, is it
> necessary for the entire probe to be phosphorothioated? Second, when adding
> oligo to tissue culture flask is there one way to combine better than
> others??? i.e. should I combine oligo and lipofectin prior to adding to cells??
> Thanks in advance.
> Karen Gravitt
When adding the oligo to culture flask you should pre-mix the oligo and
lipofectin for a while (at least 15 min), and you also need to work out
the concentration of lipofectin, and to be sure that the lipofectin only
does not kill the cells.
Hoping that may help.
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