UPLC Separation of DNA Duplexes

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Sean M. McCarthy and Martin Gilar
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Application Notes
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Besides gel electrophoresis, Waters UltraPerformance Liquid Chromatography (UPLC) Technology can be used for rapid and cost-effective separation and purification of wide array of double-stranded DNA sequences (dsDNAs).

This application outlines a UPLC method for the separation/purification of dsDNA sequences ranging from 50 to 600 base pairs in length, in under 20 minutes. The separation is based on DNA length, rather than on the sequence. Because of the substantial column mass load capacity, large amounts of dsDNA fragments can be isolated.

This presented method dramatically reduces analytical time and effort compared to gel electrophoresis, and can be utilized for separation of dsDNA fragments produced by hydrodynamic shearing. Additionally, isolated samples may be used for next-generation sequencing. With current research efforts focused on reducing total genome processing times and sequencing costs, the Waters ACQUITY UPLC System solution provides for a superior tool to those currently used for dsDNA isolation, purification and analysis.

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