I would like to do neurite outgrowth analysis and live-dead viability assays with Calcein/Ethidium Homodimer-1 on PC-12 cells. I would like to know if anybody has tried these stainings prior to RNA isolation with TRIzol reagent. I seed my cells on a dark-colored scaffold, therefore I'm planning to use Calcein-AM to make visible the neurite structures while neurite length quantifications.
I am wondering if it is possible to isolate RNA after calcein staining and if the dyes cause decrease in the RNA quality/stability or puts them under stress (so that the RNA profiles of the target gene would be disturbed). I'd be glad to get any comments. Thanks.
Dear Idil, I´m sorry I don´t have an an answer for your question but I´m asking this myself at the moment too. I stumbled up on your question when trying to find out if its possible or not :). Did you by any tested it and can let me know if RNA isolation after Calcein/EthidiumHomodimer-1 staining was successful? Thank you!
Dear both,
I am also looking for an answer to this question, so did you try this ?
Thanks alot!
Dear Sarah and Jasmin,
Eventually I did try this but the scaffold I seeded my cells on was quenching fluorescence, so it was of no use for me. Now I don;t work there anymore, but would be interested to know the answer, if someone else tried.
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