I'm going to be isolating RNA (using the Trizol Reagent method) from a bladder cancer cell line and would like to store my isolated RNA in the freezer.
I see that in the Trizol protocol, it gives 3 stages in which you can freeze your sample:
1. Placing cells in Trizol and freezing them at -20C.
2. The washing stage, where the RNA is stored in 75% ethanol at -20C.
3. A the end, where the sample is resupended in DNase/RNase-free water and frozen at -70C.
I currently cannot do option 3 as I am waiting on our order of RNase/DNase-free water to arrive. Which one would be the most suited/which would you recommend?
Amy Kinsella
Hi,
I understand your situation; instead of choosing option 2, use option 1 because option 2 requires you to recentrifuge and extract suspended RNA again. As a result, option 1 is my preference.
Regards,
Deepdarshan Urs
I agree with the others, it's preferable to store homogenised cells in Trizol at -80 rather than ethanol precipitates of RNA. I will also say that if your experiments are delayed because you don't have DNase/RNase free water, there's a very good chance you can scrounge some from a kit your lab already has. Tubes of ultrapure water are very commonly supplied with: column kits (for DNA or RNA extraction, cleanup, gel purification etc), PCR kits, reverse transcription kits, etc. Just thought I'd mention it because I know the frustration of being delayed while waiting for reagents and this should be a pretty easy one to source!
Thanks Laura Leighton !
I did end up storing the homogenised cells in Trizol at -80. What I plan to do (hopefully!) is store each experiment's homegenised cells like this and then do the RNA extraction at the end!
Unfortunately, we don't actually have DNase/RNase free water lying around as I'm the only person working in the lab at the moment!
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