What is Tris buffer for DEPC processing
Tris buffer is a buffer prepared with tris(hydroxymethyl)aminomethane Tris. DEPC treatment refers to a sterile Tris buffer that has been treated with DEPC (diethyl pyrocarbonate) and is usually used for RNA research. Used as a solvent and pH buffer for nucleic acids in experiments.
Tris buffer treated with DEPC
The role of DEPC in Tris buffer
When Tris buffer is used in RNA-related experiments, the water used in the preparation of the buffer is required to be high, and the solution may contain a small amount of RNase, which will cause the degradation of RNA. DEPC can inactivate various proteins. It is a strong but incomplete nuclease (RNase) inhibitor. It can react with the imino, sulfhydryl, hydroxyl and other groups of many enzymes, destroying the binding site of the enzyme and causing it Its inactivation. The DEPC-treated Tris buffer can prevent RNA from being degraded by nucleases.
The amount of DEPC in Tris buffer
Generally speaking, 0.1% (volume fraction) of DEPC in the aqueous solution can inactivate the nuclease Rnase, but the use in Tris buffer is different. Some people think that DEPC cannot be added to Tris buffer. Tris is an amine that will accelerate the hydrolysis of DEPC. However, this statement is not true. The concentration of DEPC in Tris buffer can be increased to 1%, so that DEPC can be combined with DEPC even if it is hydrolyzed. The 0.1% concentration has the same effect.
Prepare Tris buffer with DEPC treated water
Due to the conflict between Tris buffer and DEPC, you can also use DEPC to treat the water to inactivate the RNase, and then use the treated water to dissolve the dried Tris powder to prepare the buffer. DEPC slowly dissolves in water, begins to suspend in the solution in the form of small droplets, and gradually dissolves after stirring. DEPC decomposes into ethanol and carbon dioxide very slowly. The method of autoclaving can destroy DEPC, or the solution can be heated and boiled for more than 15 minutes.
When preparing the Tris buffer for DEPC treatment, attention should be paid to DEPC, which has potential carcinogenic risks. It should be operated under ventilated conditions as far as possible to avoid contact with skin. Desheng has rich experience in the development and production of biological buffers, and can provide Tris powder reagents with high quality and low price.
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