Hello
I am doing an experiment on fish and i want to isolate RNA from spleen. I tried different kit (qiagen, geneall) but when I check the RNA quality on TAE gel i get a degradation of RNA. you can check the RNA pic in this link https://www.researchgate.net/post/Is_the_RNA_from_my_extraction_degraded#558bc2a960614b987a8b45b5.
My question while collecting the spleen from the fish what should i do?
Hey Adham. Please make sure you are keeping your samples on ice during the extraction. RNA degrades easily.
Hi Adham,
You should take some precautions during isolating RNA from solid tissue,
1. after dissecting spleen, you have to start RNA isolation within 10-30min, in the mean time keep tissues in ice.
2. Use cleaned motor pristle (treated with 0.2N NaoH first, then autoclaved, after that trated with DEPC water).
2. Try to do RNA isolation in a isolated clean room (better to use safety cabinet).
3. Use seperate set of gloves and microtip box.
Sometime RNA degration came from gel setup (gel chamber, casting tray, comb) so these were similarly treated with o.2N NaoH for 4-6 h, then clean it properly.
I hope you will get better result.
thanks you so much for your help. i will try to isolate RNA from spleen using Trizol followed by RNEasy kit and i will let you know the results.
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