Method for quantitatively monitoring mRNA capping efficiency

• Main Authors: ROSS, RICHARD, L, DU MIN
• Format: Patent
• Language: Chinese; English
• Published: 19.03.2024
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

The present invention relates to a method for quantifying mRNA capping efficiency, the method comprising mixing a sample, an enzyme mixture comprising a non-specific single-stranded nuclease and an acid phosphatase, and an isotope standard solution in a buffer solution to produce an incubation mixture, the isotopic standard comprises isotopically labeled m7G and isotopically labeled 2 '-O-methylated nucleoside, incubating the mixture, and analyzing the mixture using liquid chromatography-mass spectrometry to determine at least one of capping efficiency and 2-O-methyltransferase efficiency. 本发明涉及一种用于定量mRNA加帽效率的方法，该方法包括将样本、酶混合物和同位素标准物溶液在缓冲溶液中混合以产生温育混合物，该酶混合物包含非特异性单链核酸酶和酸性磷酸酶，并且该同位素标准物包含同位素标记的m7G和同位素标记的2'-O-甲基化核苷；温育该混合物；以及使用液相色谱-质谱分析该混合物以测定加帽效率和2-O-甲基转移酶效率中的至少一者。