I am doing RNA-sequencing on nonhuman primate brain tissue. The brain tissue was not sectioned prior to being put into RNAlater and frozen in a minus 80 freezer, so I am wondering:
1) if I want to collect small amounts of tissue at a time, how do I avoid thawing and refreezing tissue repetitively? If the tissue is in RNAlater, will the tissue be okay thawing to about room temp (or on ice?)
2) how long do I have to weigh the tissue before the RNA begins degrading?
3) the protocol is very strict that we cannot use more than 100mg of tissue, so weighing is essential. How do you weigh without tissue thawing? Again, is it okay if it is just for a few minutes? And if it was previously stored in RNA later?
Thank you very much.
According to the manual of RNALater, thawing and refrezzing does not affect the amount or the integrity of the recoverable RNA.
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