RNase decontamination

The quality of the RNA is the most important factor for the success of array analysis, and great care should be taken to ensure top quality. It is essential to work as quickly as possible during cell disruption to denature RNAses found in tissue cells before they can degrade the sample RNA. RNAses are also found on lab benches and on hands. Therefore, RNAse decontamination of the bench area used for isolation of the RNA is recommended. This is done using RNAseZap. Gloves should be changed frequently. 

All solutions used for reverse transcription must be free of RNase. If all solutions are prepared with RNase-free water in disposable plastics and no contact with RNase-contaminated spatula or pH probes has occurred, the solutions need not be treated any further. Otherwise, to decontaminate the solutions add diethyl pyrocarbonate (DEPC) (e.g., Sigma, St. Louis, MO) to the solution to a final concentration of 0.1%, shake, let sit overnight with loosened caps, and then autoclave for 15 min. Note that DEPC might be carcinogenic and that solutions containing Tris cannot be decontaminated with DEPC, but must be prepared with special caution to prevent any RNase contamination. 

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