Hi all,
I did read few papers that write protocols with overnight incubation with isopropanol for increase RNA recuperation, but the handbook (f.e. qiazol) does not mention it, only an incubation per 10min.
I have doubts, Will it true that overnight incubation increase RNA recuperation???
Hi Luis,
I would say that it depends on the buffer your RNA is in, and the concentration. Nucleic acids are less soluble in isopropanol than in ethanol, so you will get better precipitation of low RNA concentrations with isopropanol. HOWEVER, isopropanol is also better at precipitating salt, ESPECIALLY if incubated for a long time at cold temperatures (e.g. overnight at -20).
Unless you have particularly large RNA species or low concentrations, I would try to stick to ethanol, or stick to the 10 minute, room temperature incubation with isopropanol.
If you'd like to read more, BiteSizeBio (a great techniques-focused blog) has a whole post on this topic:
http://bitesizebio.com/2839/dna-precipitation-ethanol-vs-isopropanol/
I hope that helps!
Dan
Hello,
I have just tested this long incubating protocol for isopropanol. It seems that it would finally get better RNA concentration.
Hi,
You might also try to use glycogen as an inert carrier for precipitation of nucleic acids. RNA-grade available from several vendors. This may possibly allow you to return to ethanol which - as said above - will reduce the likelihood for co-precipiation of salt.
We could reliably collect < 0,1 µg mRNA in a *visible* whitish pellet of glycogen which did not interfere in any type of reactions done afterwards
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA