Hello,
In my current project I am at present trying to tackle an issue of rapidly stabilizing bacterial RNA in a large environmental sample. Most known stabilization reagents are not applicable in my case.
Essentially, I need to be able to stabilize the cellular RNA while leaving cells intact and at a pH higher than 7+ and at low salinity (this is important for subsequent cell collection step)
I have seen studies utilizing ammonium sulfate for heavy protein precipitation, and I was wondering if it does affect the RNA too, and if does not disrupt the cells.
Thank you!
Ammonium sulfate is the major component of RNA stability kits (as RNAlater etc). Most often the supersaturated solution of ammonium sulfate (80g/100ml) is used. I have no experience with bacterial cells, but recently I have succesfully isolated RNA from rat liver stabilizated in ammonium sulfate solution (80g/100ml) with no additional components (even no buffer), which stored 7 years in freezer (-25 C).
Important to note that you should totaly remove the ammonium sulfate if you use RNA extraction technique based on Chomczynski method (by acid guanidinium thiocyanate-phenol-chloroform extraction).
Dear Artur,
don't bother much about the integrity of RNA and DNA in ammonium sulfate solutions. More important is pH of the solution and temperature above r.t. DNA is less stable at pH low 4.5 because of apurinization and RNA is hydrolyzed at pH higher than 10 at the temperature above r.t.
Ammonium sulfate is the major component of RNA stability kits (as RNAlater etc). Most often the supersaturated solution of ammonium sulfate (80g/100ml) is used. I have no experience with bacterial cells, but recently I have succesfully isolated RNA from rat liver stabilizated in ammonium sulfate solution (80g/100ml) with no additional components (even no buffer), which stored 7 years in freezer (-25 C).
Important to note that you should totaly remove the ammonium sulfate if you use RNA extraction technique based on Chomczynski method (by acid guanidinium thiocyanate-phenol-chloroform extraction).
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA