Hello everyone,
I had had a difficulty in obtaining good RNA yield from sheep pancreas samples in the past. This time I am going to try with mice samples. Would you please suggest effective ways to preserve mice pancreas to obtain good RNA yield for qPCR?
I would really appreciate for your kind cooperation in it.
you can do the following
1- try to use the RNA later (Qiagen) solution
2- keep samples at -80oC
3- during homogenization , be rapid with optimized buffer
4- use the maximum volume of elution buffer and incubate in Silica tube 1min in Room temp. before centrifugation ,then recentrifuge by the same ellute
Homogenizing the tissue directly in TRIzol immediately after dissection and brief nuclease-free PBS rinse, then freezing the TRIzol-tissue homogenates/slurries at -80C is a very good way to avoid tissue nuclease-mediated RNA degradation. [0.1 g tissue to each 1 mL TRIzol].
Then, when isolating RNA, thaw the slurries, vortex for 5 seconds, let stand for 5 minutes, and proceed with usual TRIzol procedure for RNA isolation.
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