Technique / Genetics / Cytogenetics analysis

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By Olalekan Oguntade on 20-Apr-2008 I am working with fish samples, i found it difficult fixing my samples. I injected my fish with 0.06%Colchicine (1ml/100g BW) 4hrs before crushing the gill epithelium and tissue and kidney. I then placed in 0.75%KCl for 30 minute @ room temp before centrifuge for 10minute at 3000rpm. the sediment was then fixed with glacial acetic acid and methanol (1:3) and then centrifuge before staining in 5% Giemsa solution in 6.8 Phosphate buffer for 30 minutes. Please advise and suggest better protocol. Regards Rating: n/a Reply

By ahmad on 15-Jan-2008 Could someone please advise me about my cytogenetic cultures. I am facing a situation where the blood after settling on the bottom of culture flask assumes a shape of a flake or a viscous clot. I am using standard protocols using 5 ml RPMI-1640 culture medium, 1 ml FBS, 50 micro liter of P/S, 100 mic-liter of l-glutamine and 50 micro liter of phytohaemaglutinin cultured for 72 hours. But the troubleshootin even starts after 12 hr culturing at 37c Rating: Very Good Reply