Although we worked very fast and used RNAlater according to manufacturer instructions, we have insufficient total RNA integrity (far below a RIN of 5).
We used the RNeasy Mini Kit for total RNA extraction.
Tissue and total RNA storage occurred at -80°C.
Does anyone had the same problems and found a way to solve this problem?
Any advice would be much appreciated!
Kind Regards,
Daniel.
Flash freeze the pancreas in liquid nitrogen, grind it and proceed to make lysate in Trizol or Lysis buffer from an RNA prep kit...........RNAlater doesn't help preserve RNA in pancreas in our experience too.........
So, the key factor is the time post-disection.
We have to consider this in future studies! Thank you very much for your help!
Did you ever tried this approach?:
http://www.biotechniques.com/BiotechniquesJournal/2012/May/Simplified-and-versatile-method-for-isolation-of-high-quality-RNA-from-pancreas/biotechniques-330537.html
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