How To Remove RNase or DNase From Plastic Containers
RNase, an enzyme that breaks down RNA, and DNase, which breaks down DNA, are contaminants that can interfere with nucleotide research. DNase can be destroyed by autoclaving for 15 minutes at 121C or by following any of the procedures listed below. One or more of the following techniques will inhibit or remove RNase from your plastic container. Match the resin code on the bottom of your NALGENE container with the correct technique.
Heat at 180C for at least 8 hours.
Rinse in chloroform.
Soak in a 0.1% aqueous solution of diethyl pyrocarbonate (DEPC) for 2 hours at 37C; rinse several times with sterile (DEPC-treated) water***; heat to 100C for 15 minutes OR autoclave for 15 minutes at 121C on a liquid/slow exhaust cycle. (Heating or autoclaving will remove DEPC residues.) Note heating variations in the following chart.
Clean equipment with a detergent solution, rinse thoroughly with water and rinse with 95% ethanol to dry. Soak the equipment in a 3% hydrogen peroxide (H2 O2) for 10 minutes at room temperature. Rinse the equipment thoroughly with DEPC-treated water.***
Soak equipment in 0.1N Sodium Hydroxide (NaOH) in 0.1% EDTA in water overnight and then rinse thoroughly with DEPC-treated water.
* Rinse only, no long-term contact
**Rinse copiously to minimize chemical attack
***DEPC-treated water: Add 0.1% DEPC to water and allow to sit for at least 12 hours at 37C. Then heat the water to 100C for 15 minutes or autoclave at 121C (250F) for 15 minutes.
(^1) Sambrook, J.; Fritsch, E.F.; Maniatis, T.; "Extraction and Purification of RNA''; Molecular Cloning: A Laboratory Manual, Second Edition; 7.3, Cold Spring Harbor Laboratory Press (1989)
(^2) Caution: DEPC is a suspected carcinogen and should be handled with care. DEPC solutions are irritating to the eyes, mucous membranes and skin.
(^3) Titus, David E.; Nucleic Acid Detection, Purification and Labeling; Rapid Isolation of Total RNA; PROMEGA Protocols and Applications Guide, Second Edition; pp. 125-126, 203; Promega Corporation (1991)
Information courtesy of Thermo Scientific - Nalgene