MOPS is an organic compound with molecular formula of C₇H₁₅NO₄S and molecular weight of 209.26. MOPS is an important biological buffer, which is often used to prepare RNA electrophoresis buffer. It can not only be used as a buffer system of cell culture medium, but also a common electrophoresis buffer and protein purification chromatography buffer. The patent CN114317704B introduces a method and kit for detecting N6-methyladenine in nucleic acid molecules. The MOPS in the patent is mainly used as a buffer component, this week, YACOO editor introduced a method to detect N6 methyladenine in nucleic acid molecules. The specific steps are as follows:

(1) The amino group of guanine in the nucleic acid molecule to be tested is protected to obtain the protected nucleic acid molecule;

(2) In the presence of carbonyl compounds, the protected nucleic acid molecules react with nitrite in a solvent to convert adenine in the nucleic acid molecules to hypoxanthine. The solvent is water, MES buffer, sodium acetate buffer, MOPS buffer, PIPES buffer, HEPPS buffer, TRIS buffer or any combination thereof；

(3) Optionally, deprotecting the product obtained in step (2);

(4) Detect the products obtained in the previous step.

The method of the invention uses nitrite to deamination adenine in the nucleic acid molecule to be tested and convert it into hypoxanthine, while N6 methyl adenine cannot deamination due to its stable chemical properties, so that the detection of m6A modification level or m6A site can be realized by analyzing the signal of adenine or hypoxanthine before and after transformation. This method has high sensitivity and accurate quantification, and can detect N6 methyladenine on the whole genome or whole transcriptome at the single base level.

Reference

CN114317704B Method and kit for detecting N6-methyladenine in nucleic acid molecules.