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        DEPC

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        1534

        Procurement

        • 100ml DEPC from Sigma (D5758) [1] - �33/$292 (as of 2009-01)
        • 100ml DEPC from MPI (150902) [2] - �13 (as of 2009-01)

        DEPC-treatment of solutions

        (More DEPC is needed to inactivate larger amounts of RNases, but larger amount of RNases take longer to remove subsequently and increase the risk of residual DEPC, health risk, and interference with downstream applications. 0.05% is recommended, for example, in RNA methodologies by Farrell, whereas 1% is probably the most repeated number.)

        Protocols state very different autoclave times. If the fruity smell is faint, autoclaving was sufficient. Even after a long 45min autoclave treatment the smell will not be completely gone. If you use more than 0.05% and have a very sensitive downstream experiment, it does not hurt to autoclave longer or twice.

        Warnings

        Effect of residual DEPC on RNA

        Traces of DEPC modify purine residues (A+G) in RNA by carboxymethylation. Therefore, DEPC must always be removed from solutions or containers by autoclaving or heating at 100°C for 15 min. Cell-free translation of carboxymethylated RNA will yield less protein than with unmodified template. Hybridisation is not seriously affected unless the RNA probe is heavily carboxymethylated.

        Safety

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