How to pick up stably transfected colony

Martin Houle martin.houle at REMOVE.drs.crchul.ulaval.ca
Thu Dec 10 11:12:14 EST 1998


In article <743ds7$j8n$1 at unix2.glink.net.hk>, "Andrew Leung"
<leungkc at glink.net.hk> wrote:

> Hi all,
> 
>     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.
> Since my stable transfectant cells are expected to come up within the
> following one or two days, I need someone's advice urgrely.  Thank.
> 
> Andrew
> 
> Dept Anatomy
> Chinese U, HK
> leungkc at glink.net.hk

Hi,

This is what I do to isolate colonies.

1) grow my transfectant colonies in petri dishes.

2) locate colonies on microscope and mark the underside of the dish where
I find them.

3) prepare cloning rings by cutting 1ml pipettor tips 0.75cm from the base
of the tip. autoclave

4) put some silicone vacuum grease in a small beaker and autoclave


Isolateing the colonies:

5) wash the petri a few times with PBS and leave a small amount of PBS
with cells to prevent them from drying up

6) dip the end of the cloning rings in the grease and apply them on the
colonies.

7) put some trypsin in the rings and let stand a few minutes..... 

you get the idea ....



Good luck

-- 
Martin Houle M.Sc.
Centre de Recherche du CHUL
Laval University, Quebec, Canada
email: martin.houle@(REMOVE)drs.crchul.ulaval.ca



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