How to pick up stably transfected colony
dkim at NMSU.Edu
Thu Dec 10 14:13:03 EST 1998
Another way to do this is to drain the flask and then gently wipe a colony
with a sterile Q-tip (cotton-tipped wooden rod) soaked with Trypsin-EDTA
solution. After a bit, place the cotton tip into a well or flask with
medium plus serum. Enough cells are stuck to the cotton to recover.
dkim at nmsu.edu
Ian A. York <iayork at panix.com> wrote:
: In article <743ds7$j8n$1 at unix2.glink.net.hk>,
: Andrew Leung <leungkc at glink.net.hk> wrote:
: > This is the first time I transfect cells. I just wonder in what way a
: >single colony in a culture flask could be picked up manually to proprogate.
: If you've got them in flasks, then you're going to have a hard time
: picking individual colonies by cloning rings--you need petri dishes for
: that, unless you have very delicate fine motor control and a pipetter that
: goes around corners.
: A solution that's almost as good is to do clonal dilution. Take all the
: cells from the flasks, run serial dilutions, plate them into 96-well
: plates (suitable for tissue culture, of course) at, say, 30, 10, 3 , 1,
: 0.3, 0.1 cells per well. One of those plates should have colonies growing
: in around 1 well in three. (In theory, you only need to do the 0.3
: dilution, but counting is imprecise enough that it's a good idea to do a
: Where the colonies are growing at 1/3 or less, then you can be almost
: certain that each one is a clone. Amplify it and off you go.
: Ian York (iayork at panix.com) <http://www.panix.com/~iayork/>
: "-but as he was a York, I am rather inclined to suppose him a
: very respectable Man." -Jane Austen, The History of England
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