Our lab has Tris-HCl and nuclease free water (commercially purchased), but how do I prepare this together with the nuclease free water to ensure that it doesn't degrade the RNA I am using?
No need to treat the water with DEPC when you purchased nuclease-free water and Tris-HCl. Just ware gloves, when preparing the solution. If you have the Tris as powder, don't use spoons, shake the powder directly on balance paper, dilute with your nuclease free water and adjust pH to what you needs.
Hello David
You could treat your solution with DEPC (diethyl pyrocarbonate) and boil it to decompose DEPC. DEPC is an alkylating agent and a potential carcinogen, so do this under the hood.
Good luck
Pierre
yes, DEPC is good solution to it. it is also used to resuspend RNA. but it has to be deactivated by autoclaving it. use 0.1% DEPC to prepare the buffer.
hi david , usually DEPC is used to treat the solution to make it RNase free; which was also said by pierre and sansrity. but it is ineffective on the solutions containing Tris and HEPES. so better treat your water first with DEPC, autoclave it and use it for buffer preparation
No need to treat the water with DEPC when you purchased nuclease-free water and Tris-HCl. Just ware gloves, when preparing the solution. If you have the Tris as powder, don't use spoons, shake the powder directly on balance paper, dilute with your nuclease free water and adjust pH to what you needs.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA