I have been trying to prepare root tips for chromosome counting studies in okra (Abelmoschus), but every time I am failing to get stained root tips with Feulgen stain. The root tip portion is not staining whereas the remaining portion is getting stained. Why is it happening so?
Herewith I am attaching my regular protocol for chromosome preparation. Kindly suggest me the correct protocol. Please let me know where am I doing mistake?
Try different fixarives and various concentrations of stians. Also folloq the paper mentioned above.
Should we try count Okra chromosomes under microscope ????? or any software is there.....?????
Dear Dr. Nagaraju
The Okra have 120 chromosomes and direct observation could be too time consuming and inappropriate method for this plant.
Best wishes
Dear Kattula
The root tips are collected from young seedlings between 9.30 to 10.30 a.m. and pre-treated with 0.002 M 8-hydroxyquinoline for 2-4 hours at 10-12 C and then fixed in 1:3 glacial acetic acid: alcohol for minimum of 30 minutes. Hydrolysed in 1 N HCl at 60 C. Then they are stained in Schiff’s reagent and squashed on a clean slide using 1.5% aceto-orcein stain.
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