What experiments have you used DEPC?

DEPC has been used to purge reagents of nuclease activity prior to use in an experiment involving the purification of RNA because autoclaving does not ensure the complete elimination of RNase activity; buffers treated solely by autoclaving have the potential to maintain residual RNase activity.

Why DEPC treated water is not recommended for PCR?

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.

What is the use of DEPC in RNA isolation?

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. Water is usually treated with 0.1% v/v DEPC for at least 2 hours at 37 °C and then autoclaved (at least 15 min) to inactivate traces of DEPC.

How do you prepare 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 purpose of DEPC treatment?

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 treat glassware with DEPC?

Glassware and plasticware Soak plasticware (2 hours, +37°C) in 0.1 M NaOH/1 mM EDTA (or absolute ethanol with 1% SDS), rinsed with DEPC or DMPC (dimethyl pyrocarbonate) treated water and heated to +100°C for 15 minutes in an autoclave.

Is DEPC water deionized?

Thermo Scientific deionized, diethylpyrocarbonate (DEPC) treated and 0.22 µm membrane-filtered water. It is ideal for applications involving RNA. Free of endo-, exodeoxyribonucleases, ribonucleases, and phosphatases.

What is the difference between DEPC water and nuclease-free water?

Nuclease-Free Water is prepared in a proprietary process, which yields DNase, RNase, and nuclease-free, deionized water without the use of chemical additives, such as diethylpyrocarbonate (DEPC). Nuclease-Free Water is provided in nuclease-free containers.

How do you handle DEPC?

you can put 0.1% DEPC and plastic to be treated into a sealed plastic bag. 37oC 24hr incubation is necessary. Additionally, DEPC is unstable under the light, so you should keep the bag in dark place during the incubation.

How do you prepare DEPC water or nuclease free water?

Popular Answers (1)

  1. Get MilliQ (reverse osmosis purified) water.
  2. Add 1 ml DEPC (Diethylpyrocarbonate) per 1000 ml of MilliQ or double distilled water (i.e. to a final concentration 0.1%) and mix thoroughly.
  3. Let the DEPC-mixed water incubate for 12 hours at 37°C.
  4. Autoclave DEPC-mixed water for 15 minutes.

What is the pH of DEPC water?

But what i have observed many times is that the pH of the autoclaved DEPC treated water is around 3.97-. 4.08.

How is DEPC tube treated?

It is not necessary to treat your tips and tubes with DEPC, as long as sterilization at about 200 ℃. Add 0.1% DEPC in ddH2O. Mix it with magnetic strirer for about an hour in dark. Dip all the tips and tubes completely merge in 0.1% DEPC water overnight in dark.