I have tried to isolate RNA from different cnidarian species, and I got really bad yield in one of them, and bad quality and degradation In the others (according to RIN results). I don't know if it's something in these organisms (Orders: Zoantharia and Actiniaria) or in the protocol (however I have used the same protocol with other taxa and I don't have this issue). I hope some of you can help me .
We collected some samples and they were put in seawater and transported to the laboratory. Samples were washed in distilled water and transferred to 10 volumes of RNAlater. We storage these samples at -20 C and -80C until processing. Before to add QIAZOL, samples were powdered with a porcelain mortar and pestle under liquid nitrogen.
Katherine Medina-Ortiz What RNA isolation regent are you using? TRIzol? Phenol-Chloroform extraction?
Try with phenol/chloroform method it will work out.
Here is link for optimized protocol.
Best
hope these links are useful
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2813910/
www.bioline.org.br/pdf?ej06081
https://www.thermofisher.com/.../rna-isolation/.../ten-ways-to-improv...
regards
Hello Katherine. I have successfully extracted RNA from Hydractinia with TRIzol, then adding chloroform and precipitation of RNA with isopropanol and NaCl/Na3C6H5O7. If you want to know the detailed protocol just let me know!
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