Jose I. de las Heras
jose.delasheras at hgu.mrc.ac.uk
Fri Nov 17 15:07:18 EST 2000
Penny May Avoles Kianian wrote:
> I don't know if my experience can be of help to you. We work with
> plant root tips. To synchronize them we either treat them with colchicine
> for a few hours or some researchers report using cold water to synchronize.
> Colchicine probably won't be an option since it breaks down the spindle
> fibers and stops cell division progression. The cold water approach might
> be more feasible for what you are doing. Unfortunately, I am not fimilar
> with the technique to provide additional details.
The cold water treatment (0-2 degrees) works very well for plant root
tips (and no toxic waste!). How long you treat the root tips for varies
slightly with different species. For wheat I used to treat them for
20-24 hours. However, I am not sure how "happy" fibroblasts would be at
those temperatures for long periods. Plants are much more
temperature-tolerant, as the environment changes so much.
To synchronise divisions (again this was in plants) Hydroxyurea is often
used. This involves a relatively short treatment which arrests growth
(at the end of G1?), followed by removal of the HU and growth resumes...
I can't remember what HU does to the cells, but it may be worth
Dr. Jose I. de las Heras Email: Jose.delasHeras at hgu.mrc.ac.uk
MRC Human Genetics Unit Tel: (0131) 3322471
Chromosome Biology Section ext: 2316 (office)
Western General Hospital 2311 (lab)
Edinburgh EH4 2XU Fax: (0131) 3432620
Scotland - United Kingdom
"Musha ring dum a doo dum a da, Whack for my daddy-o"
More information about the Methods