Currently I am performing FISH in situ hybridization of human telomeres with PNA probes. However, one thing happened during my experiment that there is too much background noise when I observe it under fluorescence microscope. In my experiment, I used Roche blocking reagent for ISH already and I also tried to very harsh washing condition such as Formamide 70% for 2 times, 15 min each before washing with TBST 3 times, 5 min each, but the high background noise still appears on my slide.
I'm speculating that the PNA probe aggregates has lead to this result, but I don't know how to fix it. Could anyone please suggest me what could be the other possible causes of this background noise and how to solve it? Thank you very much :)
Dear
Did You use gloves with talcum? I have seen similar image because i used it.
Luis
luis alberto mendez-rosado Interesting!
Is that documented or reported somewhere?
No, i have been doing FISH for several years, it is My experience
Luis
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