Hints for RNase-free Handling
RNA is prone to degradation due its free 2'-hydroxy-group. Main causes for degradation are the activity of RNases and prolonged incubation in alkaline solutions.
Some simple precautions help to keep your precious RNA intact:
- Work always with fresh, disposable plastic consumables (e.g. mark a bag of tubes and put it aside, never grab into the bag, rather drop the tubes out of the bag. Same is true for the pipetman-tips). If you must use glassware be aware that RNases can survive autoclaving - in this case bake your glassware at 250 degrees Celcius for at least 4 hours.
- Always wear gloves (RNases are ubiquitous). Change gloves frequently (be aware that any surface you might touch with your gloves may have already been touched by a person not wearing gloves).
- Use RNase-free water/buffers. Most commercially available water is in fact RNase-free. Alternatively, you may produce DEPC-water. DEPC (Diethylpyrocarbonate) reacts with the primary amine-groups and therefore inactivates RNases but can not be used for e.g. Tris-buffers.
Strictly applying these 3 points will work in 99% of the cases. If problems persist, Maniatis et. al. recommend to clean gel tanks, benches with special solutions.
You may also contact us if you would like to discuss RNase-free handling procedures.
