I am working on RNA in situ hybridization in plant buds. I collected I sample and fixed it in the FAA liquid at 4 C, but I didn't finish for the RNA probe. Does anyone know how could I do to keep the RNA from degradation too much?
I hear from someone that I can make the cryosection firstly, store the sample at -80C, and recover by 4% paraformaldehyde before proceeding with ISH. I think this method would be feasible. How long could I store the sample?
This should be okay for some weeks to maybe half a year.
Best
Torsten
You could either make an OCT block and store it in -80C or you could embed it in a paraffin block after fixation. I have done in situ on both types of sections few months after preparing these blocks and had no problems with either
Tissues will have better signal if you fix it in FAA only for 2-4 hours, then go on with the dehydration and embedding. Embedded tissue should be ok in the fridge (4C) for many months.
If you stored it in FAA, you can toss this tissue (or use it for other analysis). You probably lost all the signal after 24 hours in FAA.
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