Good day. I have an important question about RNA degradation: I stored RNA samples in 70% ethanol with nuclease-free water at -80 for 10 months. A few days ago, the ultrafreezer was accidentally disconnected and the temperature slowly increased to -5 °C in 2 days. When I noticed, I put the samples in another ultrafreezer. I am worry about RNA degradation because I was going to use those samples for real time PCR. Could anybody give me some suggestions? I´ve read that the samples integrity should be fine but I´m not absolutely sure. Thanks for your attention and your time. I wish everybody a good day.
Hi Jorge,
Few suggestions try to keep RNA samples in RNA storage solution,secondly RNA has a shelf life of few months after that it will no longer to be uses.
Try to check quality of your RNA by running on a gel or in a agilent Bioanalyzer and then do a cDNA synthesis.
Hi Jorge, in 70% EtOH RNA is safe even at room temperature for long time, it is in fact the best way to preserve RNA. If you will find it is not of good quality, it will be due to the original preparation, not to the storing.
If it is total RNA, also a quick run (1-2 cm) on a 1.5% standard agarose gel will give you an idea of its quality, you will easily see the ribosomal RNA bands and measure the intensity ratio between the two larger ones (depends a bit on the organism you work with, since their length changes, but in general the 28S to 18S intensity ratio should be around 2:1).
Good luck!
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