Extraction of viral RNA from cell supernatants or from plasma/serum is supposed to be possible using TRIzol or TRIsure, provided that glycogen is added.
What is your own experience? I want to extract small quantities of SARS-CoV-2 RNA for PCR purposes.
Trizol is based on "The single step method of RNA isolation" that we developed over 40 years ago at the NCI-NIH. Viral RNA can be easily extracted in any lab of the world!
Hi Lionel Berthoux ,
TRIzol method is a very good method for viral RNA purification, and I have used this method successfully for a number of viruses from different sources (ranging from insects to plants).
Regarding Guido Cordoni
s concern, you have to ensure complete removal of the TRI reagent before Isopropanol step and to wash the pellet properly with 70-75% ethanol to rule out downstream interference with enzymes.Best wishes,
SD
Greetings,
Gradient ultracentrifugation, even 20% cushio, would for sure help You to rise the concentration and purity of viral RNA, avoiding host genetic material. After that, TRIzol should work perfectly.
I have used Trizol for viral RNA (HCV) extraction from plasma and PBMC and it works well even without addition of glycogen. I think with addition of glycol it will be of better efficacy.
Trizol is based on "The single step method of RNA isolation" that we developed over 40 years ago at the NCI-NIH. Viral RNA can be easily extracted in any lab of the world!
Thanks a lot Prof @ Nicholetta Sacchi for discovering this simple and efficient method of RNA purification. It has made our work with RNA much much easier.
Regards,
SD
We published a pre-print comparing TRI Reagent to automated systems with regard to purifying SARS-CoV-2 RNA for diagnostic testing
https://www.medrxiv.org/content/10.1101/2020.05.26.20099440v1
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