Cloning with FACS sorter

terrydp at globalnet.co.uk terrydp at globalnet.co.uk
Thu Mar 4 10:02:52 EST 1999


On Tue, 02 Mar 1999 11:15:38 -0400, "Chidi"
<ngwu at helix.mgh.harvard.edu> wrote:

>Hi,
>
>Has anyone tried subcloning hybridomas with a FACS sorter? I want to get
>away from manual single cell picks and serial dilutions. It will be nice to
>throw in recently fused cells (B cells to Myeloma fusions) and sort them
>into singles.
>--
>Chidi
>
>-- Do unto others as you will have them do unto you --

Dear 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

about the best single subject-specific  resource I know of in any of
the scientific areas I've scanned for on the Internet.

Best Wishes

Terry Prospero

terryprospero at hotmail.com



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