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Method for chromosome coloring


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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