2 replies to this topic

[cells]

Hi, I was wondering if there is any drawback to isolating RNA from cells frozen in -80 for a month. I had treated cells with a compound, collected the pellet, rinsed with PBS and stored the cells at -80...it's been a month and I would like to extract RNA for gene expression analysis from these cells. Will storing my cells for a month affect my RNA isolation and gene expression results? Just curious since I really don't want to retreat my cells with the compound.

Thanks.

[suz55tbird]

I often isolate total RNA from cells very successfully.  The way I do this would take exactly the same amount of time that you spent in rinsing and freezing your cells, but would result in beautifully intact RNA.  I fear that RNA from cells stored as you described would be severely compromised by degradation.
Next time:  after treating your cells, rinse well with PBS, aspirate, add 1 mL of Trizol (for a T25 flask), scrape dissolved cells up and put in a tube, then freeze.  The Trizol will protect the RNA from degradation, and can be stored at -80 for years with no apparent ill effects.

[Trof]

Storing cells frozen on -80 is not good for RNA isolation. They need to be snap frozen and stored in liquid nitrogen or lysed with Trizol and stored in it at -80.