Hi all,
In our lab we have a lot of experience extracting RNA using Trizol.
I know (and have done this) that after taking the top phase and adding isopropanol you can leave the sample in -20c to enhance yield.
I am preparing to extract RNA from critical samples for an RNA seq experiment and I have 2 questions:
1. Does someone have a reference to the fact that the RNA doesn't degrade in these conditions at -20c? I couldn't find one.
2. I also want to add glycogen as a carrier- does anyone know if the addition of glycogen will interrupt the RNA seq protocol in someway?
Thanks!
Hello
It is not recommended to do so maybe this is the reason you could not find any references. The best temperature for keeping RNA is -80.
Hi Gidi,
just been through lots of RNA preps (Trizol) myself for RNA-seq and we have been using isopropanol precipitation in a 4C cooled centrifuge, occasionally storing samples overnight at -20C but never any longer. These two steps seem to be working equally well but better than spinning at RT.
You can read these articles for some references:
https://www.thermofisher.com/uk/en/home/references/ambion-tech-support/nuclease-enzymes/general-articles/working-with-rna.html
https://bitesizebio.com/253/the-basics-how-ethanol-precipitation-of-dna-and-rna-works/
For the second part of your question you can read;
https://www.researchgate.net/post/Could_someone_tell_me_whether_glycoblue_in_RNA_samples_will_affect_RNA_sequencing
Lots of people do it and does not affect RNA-seq.
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts