knck out strategy

Robert Mihalek rmihalek at partners.org
Wed Jun 20 07:44:55 EST 2001


"lily king" <lily.king at cshs.org> wrote in message
news:20010619231549.7435.qmail at ww02.jatek.com...
> When I inject my  DNA  into the ES cell,it is better be linearlise DNA.
> Does the DNA has to be included the whole NEO-plasmid?or it can be pop out
> the rebination DNA,like this:
>   sac11-TK-left arm-NEO-right arm-TK-sa11.
> Thanks for helping me
>
>
> <http://www.biowww.net/forum/read.php?f=1&i=2580&t=2580>
>

Dear Lily,

You should linearize the DNA prior to electroporation. Don't worry about the
plasmid sequences. In a homologous recombination event, the plasmid
sequences should not insert into the ES cell DNA. They may be present in a
non-homologous recombination event, but my guess is that you'll select
against those in culture.

Bob M.
Boston, MA






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