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74
RNeasy Mini Handbook 10/2019
Nondisposable plasticware
Nondisposable plasticware should be treated before use to ensure that it is RNAse-free.
Plasticware should be thoroughly rinsed with 0.1 M NaOH, 1 mm EDTA* followed by
RNAse-free water (see ”Solutions”, page 74). Alternatively, chloroform-resistant plasticware
can be rinsed with chloroform* to inactivate RNAses.
Glassware
Glassware should be treated before use to ensure that it is RNAse-free. Glassware used for
RNA work should be cleaned with a detergent,* thoroughly rinsed, and oven baked at 240°C
for at least 4 h (overnight, if more convenient) before use. Autoclaving alone will not fully
inactivate many RNAses. Alternatively, glassware can be treated with DEPC* (diethyl
pyrocarbonate). Fill glassware with 0.1% DEPC (0.1% in water), allow to stand overnight
(12 h) at 37°C, and then autoclave or heat to 100°C for 15 min to eliminate residual DEPC.
Electrophoresis tanks
Electrophoresis tanks should be cleaned with detergent solution (e.g., 0.5% SDS),* thoroughly
rinsed with RNAse-free water, and then rinsed with ethanol
†
and allowed to dry.
Solutions
Solutions (water and other solutions) should be treated with 0.1% DEPC. DEPC is a strong, but
not absolute, inhibitor of RNAses. It is commonly used at a concentration of 0.1% to inactivate
RNAses on glass or plasticware or to create RNAse-free solutions and water. DEPC inactivates
RNAses by covalent modification. Add 0.1 ml DEPC to 100 ml of the solution to be treated
and shake vigorously to bring the DEPC into solution. Let the solution incubate for 12 h at
37°C. Autoclave for 15 min to remove any trace of DEPC. DEPC will react with primary amines
*
When working with chemicals, always wear a suitable lab coat, disposable gloves, and protective goggles. For more
information, consult the appropriate safety data sheets (SDSs), available from the product supplier.
†
Plastics used for some electrophoresis tanks are not resistant to ethanol. Take proper care and check the supplier’s
instructions.
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