Hello Everyone!
I need to send some RNA samples to sequencing, and I want to know which RNAse inhibitor do you use to ensure that your samples don't degrade during shipping and how do you use it during the RNA isolation process.
I'll appreciate any advice
Hi Rivera-Toro,
RNase inhibitors are usually added to several enzymatic reactions such as RT, and are not generally used for RNA isolation or simple storage. You can just freeze your samples and send them in dry ice.
What I learned from my experiences is that total RNA is fairly stable once properly prepped.
Hi Diana
If your sample is precious I always advise using an RNAse inhibitor. Depending on the material and extraction method used, some RNAse could still be present. Sometimes, unfortunately, the cold chain can be broken, so it's best to use an inhibitor. I use RiboLock, it works well. Once you resuspend your final pellet, add RiboLock according to the manufacturer guidelines.
RNA degrades quickly, so although I don't completely disagree with the above experiences. I don't like to take chances.
Hope that helps
Add DEPC-treated water or DEPC directly in small quantity with RNA sample
Poly vinyl sulfonic acid is also an effective RNase inhibitor. DEPC is however, most commonly used RNase inhibitor.
DEPC should be only added to water that is going to be used for RNA experiments prior to the experiments and autocalved before the experiment to get rid of the DEPC, otherwise it will likely interfere with downstream applications or with RNA itself. Personally, I prefer to use autoclaved milli-Q water instead of DEPC water, but we do use RNAse inhibitors for some types of experiments.
Resuspending in DEPC treated water and sending in dry ice is agood idea.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA