First, it is important to set aside all reagents and equipment solely for RNA extraction and analysis. By far the most common source of exogenous RNase contamination are the hands, therefore, all manipulations of reagents and
equipment (tips, tubes, gel combs, etcetera) should be performed while wearing gloves, which should be changed
frequently. To minimise the activity of endogenous RNases that are liberated during celllysis, all manipulations should
be performed quickly, on ice where possible. RNA should be stored in small aliquots, preferably at −70◦C; repeat freezethawing of samples should be avoided. Diethyl pyrocarbonate (DEPC), a potent RNase inhibitor (Ref. 2), can be used to treat glassware and to prepare solutions (except those containing Tris buffers). However, we routinely isolate intact RNA without the use of DEPC and it should be remembered that before using any solution or equipment that has been DEPC-treated it must first be inactivated by autoclaving, thus, it no longer offers any protection against RNases.
First, it is important to set aside all reagents and equipment solely for RNA extraction and analysis. By far the most common source of exogenous RNase contamination are the hands, therefore, all manipulations of reagents andequipment (tips, tubes, gel combs, etcetera) should be performed while wearing gloves, which should be changedfrequently. To minimise the activity of endogenous RNases that are liberated during celllysis, all manipulations shouldbe performed quickly, on ice where possible. RNA should be stored in small aliquots, preferably at −70◦C; repeat freezethawing of samples should be avoided. Diethyl pyrocarbonate (DEPC), a potent RNase inhibitor (Ref. 2), can be used to treat glassware and to prepare solutions (except those containing Tris buffers). However, we routinely isolate intact RNA without the use of DEPC and it should be remembered that before using any solution or equipment that has been DEPC-treated it must first be inactivated by autoclaving, thus, it no longer offers any protection against RNases.
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