how to: picking colonies of stable transfectants

D.K. 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? 
>Thanks.
>Thomas Rasmussen
>
>http://biowww.net/mynews/tree.php?group_name=bionet_molbio_methds-reagnts&begin
>=0

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. 

DK



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