( ase contamination can effectiv...)Decontamination and inhibition of ribonucleases

ase contamination can effectively be avoided by using RNase-free solutions such as Eppendorf Molecular biology reagents and following a few common sense laboratory procedures:

• Always wear gloves when working with RNA.
• Maintain a separate area for RNA work that has an own set of pipettes, pipette tips, Eppendorf tubes, buffers, and reagents. This is especially important if your work requires the use of RNases for e.g. plasmid preparations.
• Sterile disposable plasticware produced under clean room conditions is RNase-free and should be used when possible. Eppendorf Biopur tips and tubes provide this quality feature in addition to being DNase free.
• Metal tools like spatulas can quickly be decontaminated by holding in a burner flame for several seconds. Contaminating RNases on glassware can be inactivated by baking the glassware at 180C or higher for several hours.

Alternatively, glassware can be soaked in freshly prepared 0.1% DEPC in water or ethanol for 1 hour, drained, and autoclaved (necessary to destroy any unreacted DEPC which can otherwise react with other proteins and the adenine residues of RNA). As DEPC will attack polycarbonate (e.g. centrifuge tubes) or polystyrene (e.g. standard microtiter plates) decontamination of these materials can be achieved by soaking in 3% hydrogen peroxide for 10 minutes. Residuals of pero xide can be removed by extensively rinsing with RNase-free water1., 2.. An alternative protocol uses a 1 N NaOH soak of 1 hour at 37C. After soaking in NaOH the labware is then washed extensively in RNase-free water.

RNase decontamination of buffers and solutions
DEPC treatment of solutions is accomplishe
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