Hi,
I am trying to purify synaptoneurosomes for RNA analysis from 4-week mouse brains. I am using a Percoll density gradient followed by washing 2X in buffer to remove residual Percoll (Dunkley et al., 2008). From the pellet, I have tried Trizol extraction as well as extraction using RNeasy Mini kit, and neither have been working. Does anyone have any tips or tricks for RNA extraction from synaptoneurosomes?
Thanks!
The Trizol extraction method should work which usually results in a higher yield than the RNeasy kit. Depending on the concentration and quality ratios you may have to pool samples to reach desired results. There are other tweaks that I have used (not with synaptoneurosomes) to increase yields such as freezing the samples in trizol overnight, and us a grinding pestle while sample is in trizol.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4334750/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC122371/
hope these articles might be a help. Ill have to agree with Dr. Mickey Trizol method works perfectly, the overnight trick would work indeed.
Best of Luck
Kat.
See this article light be useful: https://www.ncbi.nlm.nih.gov/pubmed/28993203
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