how to: picking colonies of stable transfectants
dk at no.email.thankstospam.net
Wed Oct 2 21:35:17 EST 2002
thoj at health.sdu.dk wrote:
>I am about to establish a stable cell line. How do I pick individual colonies
> growing on the same plate?
In addition to already suggested classic techniques of limite dilution and
cloning cylinders, here is a lazy man protocol of accimplishing the same.
Wait until clones are the size easily seen by eye, stick a yellow tip right
on a clone, scratch and wiggle a bit to detach some/most cells, suck up
~ 20 ul and transfer to 96 well plate. Chances of cross-contamination by
another clone are comparable to any other methods. Even very strongly
attaching lines can be treated this way. For people with paranoid
tendencies, the same can always be done right in the trypsin/Versen.
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