how to: picking colonies of stable transfectants

Wolfgang Schechinger hubahopp at gmx.de
Wed Oct 2 03:02:12 EST 2002


Thomas, 

I suppose you already have cells resistant against G418 resp. your
antibiotic of choice.
Then trypsinize them and seed them at a (theoretical) concentration of 1 to
3 cells per well in 96well microplate. After a few days, you should see
growing foci. Test those wells where only one focus is present for your
transgene and expand them in 12 or 6 wells.
Store the surplus cells from the first step in nitrogen for backup
purposes. 1 or 2 microplates usually are sufficient.
This method IMHO is much efficient than using cloning cylinders or whatever.

Regards,

Wo


At 20:34 01.10.2002 -0000, 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&b
egin=0
>
>

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