Molecular Weight Confirmation For Length, Integrity And Impurity Analysis Of A 70-Mer Oligonucleotide

By Remco van Soest, Kerstin Pohl, and Ling Huang, SCIEX, USA; Integrated DNA Technologies, USA

Analyze a next-generation sequencing DNA adapter using the ZenoTOF 7600 system and SCIEX OS software.

A workflow utilizing the SCIEX ZenoTOF 7600 system was used to determine the molecular weight and analyze the purity of a 70-mer DNA sequence with a 3’ dT overhang that is linked through a phosphorothioate group in the 5’… GATC*T construct.

Key features of the ZenoTOF 7600 system and SCIEX OS software for MW confirmation and impurity analysis include

• Accurate MW confirmation for length and integrity of long oligonucleotides and their impurities using high resolution TOF MS data with excellent mass accuracy and intact mass reconstruction

• Full isotope resolution after reconstruction for confident, unambiguous confirmation of MW

• Easy and accurate quantification using reconstructed mass in SCIEX OS software for quick determination of relative amounts of impurities

Review how the workflow for an NGS adapter as described in this article, can be used to analyze the length and integrity of poly(A) tails for mRNA vaccines and for single guide RNA for CRISPR content.