Hi everyone !
Does anyone have any experience in extracting RNA from exocrine pancreas (which is very high in RNAse)? The pancreas is frozen tissue. I wonder if RNAse is also high in the nucleus, if not, then possibly I could consider nuclear extraction.
Let me know your thoughts/experiences.
Thanks!
Amna
Dear Amna!
Please You look at this publication and protocol:
https://www.pancreapedia.org/sites/default/files/isolation_of_pancreatic_rna_9-15-14_pdf.pdf (attached)
Nuclear extraction is a time consuming protocol and in my experience working with brain tissue, the RNA integrity from samples after nuclear extraction is lower (around RIN 7.5) compared to processing whole tissue (RIN 9+). I don't think nuclear isolation is likely to improve your RNA quality. I think you would get more mileage out of a pancreas-specific RNA isolation protocol (someone has already linked one) or else from the standard measures for RNase-rich tissue: work fast and cold during dissection and snap-freeze your samples as soon as possible after death of the animal, homogenise frozen tissue directly in a chaotropic reagent (trizol or similar), increase the ratio of reagent to tissue.
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