RNA can be difficult to work with as it is readily degraded by RNases that are found in a variety of environmental sources, such as bacteria and fungi, as well as flaked skin and hair. RNases are extremely robust enzymes that can retain functionality at room temperature, and even after freeze/thaw cycles and autoclaving. Consequently, RNases are unaffected by many methods of decontamination, and strong chemical methods are often required to eliminate them from surfaces and solutions.
Here are a few common tips to follow to help avoid RNA degradation in your sample:
Always wear gloves when working and remember to change your gloves if you accidentally touch skin or any other contaminated surface
Use RNase-free plastics such as tubes and tips
If glassware is used, autoclaving is not enough—glassware must be baked for several hours at temperatures greater than 180°C or treated with a reagent such as one of the RNaseZap products
If preparing buffers, use water that is nuclease-free or treated with DEPC. DEPC (diethylpyrocarbonate) reacts with histidine residues of proteins and will inactivate RNases. However, it can also react with RNA, so it needs to be removed by heat treatment before the solution is used (DEPC breaks down to CO2 and ethanol).
Set up designated RNA work areas that have low traffic and are away from air vents
Protect your samples with reagents such as RNAlater Stabilization Solution and store the RNA in proper conditions:RNA is generally stable at -80° C for up to a year without degradation in RNase-free H2O with 0.1 mM EDTA or TE buffer (10 mM Tris, 1mM EDTA).
RNA solubilized in formamide may be stored at -20°C without degradation for at least one year
For long term storage, RNA samples may also be stored at -20°C as ethanol precipitates.
More information regarding RNA Preservation and Stabilization can be found in this article.