[Cell-biology] Soft agar cloning of Tumor cells

Jayakumar, R R.Jayakumar at roswellpark.org
Thu Jan 12 17:00:23 EST 2006


how about using an some enzymes like agarase to dissolve the agar.  I am not sure what effect it would have on the cells?  Does agarase dissolve agar or just agarose?
  
jai


-----Original Message-----
From: cellbiol-bounces at oat.bio.indiana.edu
[mailto:cellbiol-bounces at oat.bio.indiana.edu]On Behalf Of Ken Garson
Sent: Thursday, January 12, 2006 10:24 AM
To: cellbiol at magpie.bio.indiana.edu
Subject: [Cell-biology] Soft agar cloning of Tumor cells



Hi,

I am screening primary tumor cells for their ability to grow in soft agar.
 I have recovered adherent cell lines by sucking out the colonies with a
pipette and then mechanically breaking up the agar/cell plug to release
the tumor cells.  The mechanical disruption is not very efficient.  Is
there a method for recovering colonies from soft agar and generating a
nice single cell suspension of viable cells?

Thanks!

Ken

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