knck out strategy
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
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.
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