Cloning with FACS sorter

Chidi ngwu at helix.mgh.harvard.edu
Thu Mar 4 10:33:09 EST 1999


Terry,

Thanks for your help. It is refreshing to know I am twenty years behind, I
only wish I was twenty years younger too! Is it possible for you to email
the web page to me (I have a html mail reader)? The web page seems to be
sitting on a restricted server.

I really wonder if the sorters sort the cells sterilely, and if you have to
label the cells with a fluorescent dye? What is the viability and
conditions?

You are right, I only want to sort at about day six, after fusion and
plating in selection media.

--
Chidi



> People have been doing this for about twenty years so there's lots of
> experience.  I've done it with a BD Facstar+ and the BD single-cell
> deposition attachment.  Some hybridomas,  e.g. NS-1 based,  are more
> robust than others. Newly-fused cells are very fragile and anyway you
> want to do the usual growth on selective media BEFORE sorting to get
> rid of the large number of cells that will die.
>
> There are lots of relevant resources on the web.  Start with
>
> www.flowcyt.cyto.purdue.edu




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