I am very new to this field and am currently working on 3-color in situ hybridization with intronic and exonic probes. Surprisingly, my intronic probes that are supposed to label introns within the nucleus, falls outside of the nuclei when I see them under the confocal microscope. I am sure that my probes are specific to my gene of interest and am really confused about the results I am getting. Anybody knows why?
Is it possible 1) there are some alternative splicing variants that do in fact keep the intron of interest in the processed mRNA? Or 2) is your probe is binding nonspecifically in the cells? Maybe you could try incubating the cells with the probe at lower concentration, or with a more rigorous washing protocol.
I have just come from the EMBO congress (RNA localization and local translation) and there's people looking at the role of introns outside the nucleus. For example, the role of intron containing RNAs in dendrites, it seems that there are intron desequilibrium in synaptic genes. I don't know if it helps!
This papers says about internationalist of synthesized porphyrins and Hoechst staining in candida cells
https://www.researchgate.net/publication/279459630_Axial_Ligand_Modified_High_Valent_Tin%28IV%29_Porphyrins_Synthesis_Structure_Photophysical_Studies_and_Photodynamic_Antimicrobial_Activities_on_Candida_Albicans
Article Axial Ligand Modified High Valent Tin(IV) Porphyrins: Synthe...
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