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Abstract
An efficient one-step digestion approach generates 5'- and 3'-end fragments for mRNA analysis.
- Tailored 10-23-type DNAzymes cleave mRNA to produce both 5'-capped and uncapped short fragments alongside 3'-poly(A) tail products.
- Polyacrylamide gel electrophoresis and ion pair reversed-phase liquid chromatography analyses confirmed the successful generation of cleavage fragments in a single step.
- LC-mass spectrometry validated the production of 5'- and 3'-cleavage fragments.
- The DNAzyme-mediated method allows for direct and simultaneous analysis of capping efficiency and poly(A) tail length.
- This approach eliminates the need for additional purification steps and costly mass spectrometry, streamlining sample preparation for quality control testing.
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