Preparation of Mitotic and Meiotic Metaphase Chromosomes from Young Leaves and Flower Buds of Coccinia grandis

Materials and Reagents

1. Blotting papers
   
2. Glass slides and cover-slips
   
3. Specimen tube
   
4. 1^st, 2^nd and 3^rd leaf from the shoot apical region for mitotic study
   Note: Young leaves can be harvested from seedlings as well as from mature plants.
   
5. Flower buds of 7 and 8 stages for meiotic study
   Note: Assigned according to the length of flower buds.
   
6. Saturated solution of para-dichlorobenzene (Sigma-Aldrich, catalog number: D56829 )
   Note: It is also named “1,4-Dichlorobenzene” on Sigma-Aldrich website.
   
7. Glacial acetic acid (SRL, catalog number: 85801 )
   
8. Hydrochloric Acid (HCl)
   
9. Orcein (Sigma-Aldrich, catalog number: O7380 )
   
10. Carmine (Sigma-Aldrich, catalog number: C1022 )
    
11. Absolute ethanol (99%) (Merck Millipore Corporation, catalog number: 100983 )
    
12. 2% w/v aceto-orcein stain (see Recipes)
    
13. 1% w/v aceto-carmine stain (see Recipes)
    

Equipment

1. Compound microscope (ZEISS, model: Primo-star )
   
2. Refrigerator (household refrigerator having a freezer compartment with sub-zero temperature)
   

Procedure