This document describes the collection of fractions from an oligonucleotide separation using gel electrophoresis and offline MALDI-TOF analysis. By using the Agilent oligonucleotide kit and Agilent Capillary Electrophoresis system, antisense oligonucleotides can be successfully separated. The Agilent ChemStation software allows for fully automated collection of fractions using signal A as the peak sensor. This method enables further analysis of the separated peaks and determination of the molecular mass.
This document details an application for separating antisense oligonucleotides using a sieving-polymer-filled capillary and collecting specific fractions for further analysis. The process utilizes the Agilent Capillary Electrophoresis system with ChemStation software to automate fraction collection based on peak detection.
Following separation, the collected fractions are subjected to offline MALDI-TOF mass spectrometry analysis. The text provides specific experimental conditions, including buffer types, voltage settings, and detection parameters, demonstrating the successful determination of analyte mass.
| Capillary length | 24.5 cm |
|---|---|
| Capillary inner diameter | 100 µm |
| Injection time | 7s |
| Run voltage | -25 kV |
| Detection wavelength | 260 nm/8 nm |
| Laser energy | 4 µJ |
| Polarity | negative |
Agilent ChemStation software enables the fully automated collection of fractions using signal A as the peak sensor.
The determined mass of the collected fraction was 3230.10 Da.