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Updated: Jun 22, 2026

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
Application of a coupled reducing system to Edman sequencing
1California Institute of Technology, The Beckman Institute, Pasadena, CA 91125, USA.
Abstract:
Addition of low levels of solid tris-(2-carboxyethyl)-phosphine hydrochloride to Applied Biosystems' R4A (2 mM) and R5 (0.17 mM) resulted in a coupled, phosphinedithiothreitol reducing system. Coupling the two reductants eliminated or greatly decreased interference from the oxidation product of dithiothreitol and allowed quantitation of phenylthiohydantoin (PTH)-Asp below the 400-fmol level. Use of the system resulted in a measurable increase in repetitive yield for beta-lactoglobulin, and instrument PTH standards were stable for more than 3 months.
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