Method for chromosome coloring
- To: hosta-open@mallorn.com
- Subject: Method for chromosome coloring
- From: z*
- Date: Mon, 20 Nov 2000 11:20:00 +0100
For those of you who want to try it is here a method I have used for
students for years Remember this is done with very favorable
material ie thick roots, big chromosomes and a small number of
chromosomes Like in Vicia faba = broadbean
1.Take an actively growing root tip
2.fix it in Carnoy ( ethanol : acetic acid : chloroform 6:1:3) for 1h
3. bring the root via 60 %, 40% and 20% ethanol in water
4.Heat the root for exactly 12 min at 60 C in 1N HCl
5.rinse in distilled water
6.incubate in Feulgen reagens ( Sigma) for 1-2 in the dark
7. Wash at least three times in 400 ml water containing 2 gram
Potasssium metabisulfite ( SO2 water)
8.wash root for 30min in running tap water
9. Crush the root in 45 % acetic acid under a cover slide on a glas
10. Look for cells in mitosis under the microscope and make
drawings of all the different stages. If you cannot find any
cells in mitosis, take a new root .
I realize this is a laborious procedure Those of you who want to do
it in 30min should ask somebody else..... Or order ready-made
slides of Vicia faba from Carolina Biological supply company..
Ben J.M.Zonneveld
Clusius lab pobox 9505
2300 RA Leiden
The Netherlands
mintemp-16C(5F)
Zonneveld@RULbim.LeidenUniv.NL
Fax: 31-71-5274999
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