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Related Concept Videos

RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...

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Related Experiment Video

Updated: May 29, 2026

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications
05:41

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications

Published on: July 10, 2020

Transcript-independent quantitative analysis of RNA cap structures by LC-MS/MS.

Ryan Kilburn1, Rachel C Gomez1, Steven Evans1

  • 1Department of Research and Development, Nature's Toolbox Inc, Rio Rancho, NM, USA.

Molecular Therapy. Advances
|May 28, 2026
PubMed
Summary

A new LC-MS/MS assay accurately quantifies RNA 5' cap incorporation, a crucial quality attribute for RNA therapeutics. This transcript-independent method supports process development and quality control for improved RNA drug manufacturing.

Keywords:
LC-MS/MSRNARNA cappingcritical quality attributeexternal calibrationmRNAmanufacturing quality controlnuclease P1quantitative analysis

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Last Updated: May 29, 2026

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications
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Characterizing RNA Modifications in Single Neurons Using Mass Spectrometry
08:45

Characterizing RNA Modifications in Single Neurons Using Mass Spectrometry

Published on: April 21, 2022

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Accurate assessment of 5' cap incorporation is vital for RNA therapeutic quality.
  • Current methods are often transcript-specific, limiting throughput and broad applicability.

Purpose of the Study:

  • To develop a quantitative, transcript-independent LC-MS/MS assay for RNA cap incorporation.
  • To support RNA therapeutic process development and quality control.

Main Methods:

  • Utilized nuclease P1 digestion and external calibration with cap analog standards.
  • Employed paired digested and undigested analyses for reagent correction.
  • LC-MS/MS assay with a 10-minute acquisition time.

Main Results:

  • Demonstrated excellent linearity (r² > 0.995), accuracy (84%-102% recovery), and precision (RSD <15%).
  • Assay supports a 0.01-20 pmol calibration range.
  • Validated for multiple RNA transcripts, showing sequence-agnostic quantification.

Conclusions:

  • Developed a rigorously characterized, sequence-agnostic analytical platform for RNA cap analysis.
  • Method is compatible with chemical and enzymatic capping strategies.
  • Enables scalable process development, lot comparability, and manufacturing quality control for RNA therapeutics.