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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.(^1)
-
Rinse in chloroform.(^1)
-
Soak in a 0.1% aqueous solution of diethyl pyrocarbonate(^2) (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.(^3)
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RNase Removal Chart - Techniques
|
| Plastic |
1 |
2 |
3 |
4 |
5 |
| Resin |
(Heat) |
(Rinse) |
(Soak) |
(Soak) |
(Soak) |
Comments |
| ETFE |
|
X |
X |
X |
|
| FEP |
X |
X |
X |
X |
X |
| HDPE |
|
X* |
X |
X |
X |
Heat to 100C for 20 minutes |
| LDPE |
|
X* |
X |
X |
X |
Heat to 70C for 120 minutes |
| PC |
|
X* |
X* |
|
|
|
| PETG |
|
|
X |
X |
X |
Heat to 60C overnight |
| PFA |
X |
X |
X |
X |
X |
|
| PP/PPCO |
|
X* |
X |
X |
X |
|
| PMP |
|
X* |
X |
X |
X |
|
| Barex |
|
|
|
X |
X |
|
| HIPS |
|
|
X |
X |
X |
Heat to 70C for 120 minutes |
| PVC |
|
|
X |
X |
X |
Heat to 60C overnight |
| TPE |
|
|
X |
X |
X |
|
|
|
* 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)
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