I have been trying to isolate total RNA from healthy rat sciatic nerve but the concentration has always been very low in the range from 50-65ng/ul. I have tried different weights of the nerve, took close to 1cm of the nerve and incubated it in RNA Later at 4C for max of 3 hours and proceeded with RNA isolation immediately.
I used TissueLyser LT for homogenizing the tissue in TRIzol with a 5mm bead for 20-25 mins. And I followed the instructions on the Qiagen's RNeasy Plus Universal Mini kit to isolate the RNA. There are some additional steps that I followed like adding the RNase Free water directly to the center of the spin column and waited for 3 mins before eluting the RNA and completely drying the spin coulmn off ethanol before elution.
Can someone guide me with the issue? Or if isolating miRNA would be a good option in this case as according to this paper? http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3172250/pdf/pone.0024612.pdf
Thanks
Can I ask: What is your targeted conc?
the paper you ref. used mirVana Kit. Have you tried it?
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