Sep 20, 2023
Preparation of DEPC-treated water
This protocol is a draft, published without a DOI.
- 1Center for Molecular Biology of Heidelberg University (ZMBH), DKFZ-ZMBH-Alliance, 69120 Heidelberg, Germany.
Protocol Citation: Carlos Costas Insua 2023. Preparation of DEPC-treated water. protocols.io https://protocols.io/view/preparation-of-depc-treated-water-cz9qx95w
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: September 20, 2023
Last Modified: September 20, 2023
Protocol Integer ID: 88080
Abstract
Diethyl pyrocarbonate (DEPC) is a wide-spectrum RNase inhibitor. DEPC-treated water is generally used when working with RNA
Materials
Diethyl pyrocarbonate (Sigma-Aldrich #D5758)
Safety warnings
Trace amounts of DEPC will modify purine residues in RNA by carboxymethylation. Carboxymethylated RNA is translated with very low efficiency in cell-free systems. However, its ability to form DNA:RNA or RNA:RNA hybrids is not seriously affected unless a large fraction of the purine residues have been modified. Residual DEPC must always be eliminated from solutions or vessels by autoclaving or heating to 100°C for 15 minutes.
Add 1 mL of fresh DEPC to 1 L of H2O. Shake well to disperse the DEPC through the H2O. Incubate at 37°C for at least 12 h and/or autoclave at 15 psi on liquid cycle for 20 min to inactivate the remaining DEPC.
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