Besides obtaining the brain for RNA extraction, I need to obtain other organs in the mouse, such as the spleen, to measure neurotransmitters using HPLC (neurotransmitters degrade very rapidly!). Therefore, my question is, should I prioritize the brain and obtain it before peripheral organs, or vice-versa? And also, I would really like to know if someone is familiar with the upper time limit (seconds/minutes) of leaving unirrigated brain at room temperature, where RNA degradation advances to the point of jeopardizing RNA extraction of whole brain samples.
some studies have suggested that 2-6h is OK but I things that might be pushing it I would suggest having 2 people so it speed things up I would aim for 15mins, it is afer
Thanks a lot for your response Kevin. Do you have a link to the studies that you mentioned?
could try thisArticle Gene expression profiling in the post-mortem human brain - N...
But I would cut the mouse head off give it to a "friend" and while he/ she is taking out the brain you can do the other despite evidence that delay isnt that important I liked to get on with it
organs.https://www.nature.com/articles/3880451
and
Article Postmortem interval effect on RNA and gene expression in hum...
and
Article Postmortem Delay Has Minimal Effect on Brain RNA Integrity
Kev
I have heard of people leaving it especially with human samples but if you are doing it with mice then bring a friend, its simpler
Cheers
Kev
Hello,
In appropriate storage conditions you can store 3-6 months. Without doing freeze and thaw condition. As soon as dissect the brain in the ice cold plate transfer to the liquid nitrogen.
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