Digested mRNA Analysis of Sequence, 5’ Cap, and PolyA Tail
Comprehensive mRNA Characterization: Sequence, Cap Structures, and PolyA Tails in One Workflow
Webinar Summary
Discover a unified approach to mRNA PQA analysis with LC-MS/MS and advanced data processing in Byos. This webinar will showcase how enzymatic digestion, combined with ion-pair reversed-phase separation, enables in-depth profiling of mRNA sequence, 5’ cap structures, and PolyA tails. Using real-world data from eGFP mRNA, we’ll demonstrate how to tackle isomer resolution, quantify capping efficiency, and deconvolute PolyA length variants—all within a streamlined, high-coverage workflow.
Key Learning Points
- Learn how digested oligonucleotide mapping provides >99% sequence coverage for mRNA, even in the presence of isomeric oligos.
- Understand how to quantify 5’ cap variants and intermediates using Byos software and robust MS/MS scoring techniques.
- See how PolyA tail deconvolution and mass matching reveal length distributions with LC-resolved detail for high-confidence PQA assessment.
From The Webinar
On software capabilities: "state of the art software, ProteinMetrics, for digested oligos and also for intact as well. This software offers a rapid and accurate identification of small and also the very large oligonucleotides." 10:01
On workflow features: "This is what this software includes, MSMS or legal spectrum matching or OSM using a repurposed version of Byonic... It detects and localizes modifications. It's useful for the five prime cap identification, addition of methylation modifications to the sequence." 17:26
On overall workflow value: "Here on the right is your workflow and time digest LC MS to data processing. Here on the left is what this workflow can tell you. You can give your sequence identification of your mRNA, including isomer identification, percent cap efficiency of the five prime cap, and also poly a tail molecular weight and distribution." 31:45
On manual quality control: "The software allows you to manually click on each dot and compares course and assess the quality of the permutation to avoid any false positives." 28:19