This presentation contains the principle, reagents and procedure for viewing chromosome and it's Metaphase state.Hope this ppt help for your reference.

1. PREPARATION OF
METAPHASE CHROMOSOME
FROM ONION CELLS
By
K. VISALI
2ND M.Sc Biotechnology
Annamalai University

2. MITOSIS
 Two identical cells- same number of chromosome
 Stages:
P-prophase
M- metaphase
A- anaphase
T- telophase

3. METAPHASE
 Second step
 Alignment of duplicated chromosome
 Karyotype – find chromosomal abnormalities
 Colchicine – arrest cell division at metaphase

4.  Mitosis process visible in
 Somatic Cells
 Eukaryotic Cells
 Asexually reproducing Cells

5. Why onion used?
 MeristamatIc cells
 Monocot plant-large chromosome
 Mitosis varies
 Influenced

6. Reagent uses
 Sciff’s reagent- cytological study
 Fuchrin- dye to stain nuclear
 Potassium metabisulphite- slow’s down mitosis process
 HCl- breakdown the pectin
 Decolouring charcol- to remove coloured impurities

7. Protocol
Onion on tile
Grow root tips
Take the newly grown roots

8. Cont….
Add glacial acetic acid : ethanol (1:3)
1 day incubation
Place on slide
Add 1 drop of 1N / 10 HCl

9. Cont….
2-3 drop of acetocarmine stain
Destain

10. Observation
 Different phases of mitosis observed.

11.  THANK YOU