We have been using the Miltenyi magnetic separator for isolating neurons, astrocytes, and endothelial cells from mouse brains, but we are having problems.
We are getting a small pellet with cells when when finish the isolation. We would like to use this for PCR, so we need to further process these cells into mRNA. We have been using the RNAqueous 4 PCR kit and the Qiagen AllPrep kits. I know there is BSA in the pellet, so we wash the pellet first with PBS to remove the BSA, so it should just be pure cells going into the kit. Thus far, we haven't gotten any yields.
Any help on this topic would be greatly appreciated. Thanks!
hope this link is useful
https://www.thermofisher.com/...isolation/.../isolation-of-total-rna-fro...
regards
I would suggest to use Trireagent method to isolate total RNA. Just lyse your cell pellet using 1ml of Trireagent and isolate Total RNA from cells using Molecular Research Center's protocol. Very easy, and straight forward protocol to follow for the very high quality RNA.
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