Method for quantitative monitoring mRNA capping efficiency
AssigneeThermo Finnigan LLC
InventorsRobert L. Ross, Min Du
Life of this patent
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Abstract
A method for quantifying mRNA capping efficiency includes combining a sample, an enzyme mixture, and an isotopic standard solution in a buffer solution to create an incubation mixture, the enzyme mixture including a non-specific, single-stranded nuclease and an acid phosphatase and the isotopic standard including 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 a capping efficiency and a 2-O-methyltransferase efficiency.
Claims24
A method for quantifying mRNA capping efficiency comprising:
- combining an mRNA sample, an enzyme mixture, and an isotopic standard solution in a buffer solution to create an incubation mixture, the enzyme mixture including a non-specific, single-stranded nuclease and an acid phosphatase and the isotopic standard including isotopically labeled m7G and isotopically labeled 2′-0-methylated nucleoside (Am, Gm, Cm, or Um);
- incubating the mixture;
- and
- analyzing the mixture using liquid chromatography-mass spectrometry to determine at least one of a capping efficiency and a 2′-0-methyltransferase efficiency.
Dependencies are inferred from the claim text.