How do you use DEPC?

If you have ever worked with RNA, you know about DEPC (diethylpyrocarbonate). You add it to water at a concentration of 0.1%, shake or stir, incubate at 37°C for two hours or at room temperature overnight and, as if targeted by a magic bullet, the RNAses that may have been in the water are gone.

How do you make DEPC?

DEPC Treated Water Recipe

  1. Add 1ml of 0.1% Diethylpyrocarbonate (DEPC) to 1000ml distilled water.
  2. Mix well and let set at room temperature for 1 hour.
  3. Autoclave.
  4. Let cool to room temperature prior to use.

What is the pH of DEPC water?

Nuclease Free Water (DEPC Treated) pH: < 7.0 Storage: Room Temp.

What does DEPC water do in PCR?

DEPC-treated Water – DEPC destroys enzymatic activity by modifying -NH2, -SH and -OH groups in RNases and other proteins. DEPC treatment is a very effective way to treat solutions that will contact RNA.

Does DEPC expire?

Once opened, the bottle should not be kept for more than 9 months. 2. To decontaminate glassware of RNase, the glassware may be incubated with a DEPC solution for 30-60 minutes, and subsequently autoclaved to decompose the DEPC.

What is DEPC used for?

DEPC is a potent, non-specific chemical inhibitor of RNase 2 and its most common application has been to make water nuclease-free. DEPC is also carcinogenic and should be handled with care and with strict attention to the safety recommendations of the manufacturer.

What is the IUPAC name for DEPC?

Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).?) Diethyl pyrocarbonate ( DEPC ), also called diethyl dicarbonate ( IUPAC name), is used in the laboratory to inactivate RNase enzymes in water and on laboratory utensils.

What is the function of DEPC in RNA extraction?

It does so by the covalent modification of histidine (most strongly), lysine, cysteine, and tyrosine residues. DEPC-treated (and therefore RNase-free) water is used in handling of RNA in the laboratory to reduce the risk of RNA being degraded by RNases.

What are the alternative approaches to DEPC removal?

Alternative approaches include (1) maintaining certain solutions at 60° overnight 4, a process that should be carried out in a chemical fume hood with the bottle cap slightly loosened; and (2) removing DEPC from large volumes of DEPC-treated water by boiling it for 1 h in a fume hood.

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