How do you make DEPC water?

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.

How do you handle DEPC?

Rinse tanks with DEPC (diethyl pyrocarbonate)-treated and autoclaved water before use (see below). Treat glassware and plasticware with RNase-inactivating agents. Glassware should be baked at +180°C for at least 4 hours. Note, however, that autoclaving alone is not sufficient to eliminate RNases from your experiments.

What is DEPC water?

Diethyl pyrocarbonate (DEPC), also called diethyl dicarbonate (IUPAC name), is used in the laboratory to inactivate RNase enzymes in water and on laboratory utensils.

Is DEPC treated water sterile?

DEPC-treated water is autoclaved pre- and post-packaging to ensure sterility and inactivation of DEPC. It is rigorously tested for contaminating nonspecific endonuclease, exonuclease, and RNase activity.

How do you store DEPC treated water?

DEPC treated water can be acidic which may cause depurination of the oligo resulting in degradation. For optimal stability, oligos that are to be stored long term should be stored frozen, at–20°C.

Does DEPC treated water expire?

DEPC-Treated Water is shipped on dry/blue ice. On arrival store at -20°C for optimum stability. When stored under the recommended conditions and handled correctly, full activity is retained until the expiry date on the outer box label.

Can I use DEPC treated water for Qpcr?

DEPC treatment would be preferred for enhance sensitivity and completely remove any potential for RNA degradation by RNases during cDNA synthesis, but nuclease free water should be sufficient for PCR. However, DEPC treatment will not affect your PCR, if you choose to use this, as long as the DEPC has been inactivated.

When should you not use DEPC water?

DEPC is not recommended as it can be slightly acidic, which can lead to depurination and degradation of the oligos. Ideally a low TE buffer should be used ( 10 mM Tris pH 8.0, 0.1 mM EDTA ).

Does DEPC-treated water expire?

Is DEPC-treated water nuclease-free?

Ambion® DEPC-treated water is certified nuclease-free and supplied in five bottles containing 100 mL each. DEPC-treated water is autoclaved pre- and post-packaging to ensure sterility and inactivation of DEPC. It is rigorously tested for contaminating nonspecific endonuclease, exonuclease, and RNase activity.

Can I use DEPC treated water for PCR?

How do you mix DEPC and water?

Mix 1 mL of DEPC (diethyl pyrocarbonate) with 1000 mL of water in a screw-cap glass bottle. Incubate for ~2 h at room temperature in a fume hood with occasional swirling. Autoclave. Store at room temperature for up to 12 mo.

How does DEPC work in water treatment?

DEPC Water Treatment (OpenWetWare) Diethyl pyrocarbonate (DEPC) is an efficient, nonspecific inhibitor of RNases. It is typically used to treat water and solutions before working with easily degraded RNA. DEPC reacts with amine, hydroxy and thiol groups of proteins thereby inactivating RNAses (and other enzymes).

How do you make depc-h2o?

DEPC-H 2 O. Mix 1 mL of DEPC (diethyl pyrocarbonate) with 1000 mL of water in a screw-cap glass bottle. Incubate for ~2 h at room temperature in a fume hood with occasional swirling. Autoclave. Store at room temperature for up to 12 mo. « Previous | Next Article » Table of Contents.

How do you make diethylpyrocarbonate?

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