Gilson HPLC系统对肠杆菌噬菌体DNA切片的高分辨率纯化

Data in this application note were provided by M. Kawakatsu and Y. Hayashi, Applied
Technical Department, M&S Instruments, Inc., Japan.

As the size‐selective separation of nucleic acids is a necessary step in many molecular
biological analyses performed in clinical, forensic, and other types of laboratories, the
development of optimal DNA separation methods is important. Liquid
chromatographic separation techniques involve simple automated sample purification
steps and provide high resolution of DNA molecules, thereby enabling efficient DNA
separation. Furthermore, fragments can be identified and quantified directly when
using liquid chromatography methods.

The separation and characterization of modified DNA is particularly relevant, as
general interest in next‐generation sequencing (NGS) is expanding.1 This application
note describes the successful separation of methylated lambda phage DNA fragments
on a Gilson HPLC system using a Sepax PolyRP‐1000 column. Prior to HPLC analysis, a
gradient method was developed to provide sufficient resolution of the DNA fragments
(25‐2000 bp). These results demonstrate the feasibility of separating DNA fragments,
including those containing methylated modifications, by HPLC. This highly effective
separation method can thus be applied in a wide variety of analyses, including the
preparation steps of next‐generation sequencing workflows.