Comparison of techniques for quantification of next-generation sequencing libraries

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Abstract

To ensure efficient sequencing, the DNA of next-generation sequencing (NGS) libraries must be quantified correctly. Therefore, an accurate, sensitive and stable method for DNA quantification is crucial. In this study, seven different methods for DNA quantification were compared to each other by quantifying NGS libraries for the Ion Torrent™ and Illumina® platforms as well as dsDNA oligos with known DNA concentrations. Rather large variations in library concentration estimates were observed. The differences between the highest and lowest concentration estimates varied with a factor of 5–100 depending on the library concentration. The Bioanalyzer, TapeStation and Qubit® instruments gave concentrations closest to the expected when quantifying dsDNA oligos. At very low concentrations (2–4 pg/ul) only the Bioanalyzer could reliably quantify the dsDNA oligos.

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Hussing, C., Kampmann, M. L., Mogensen, H. S., Børsting, C., & Morling, N. (2015). Comparison of techniques for quantification of next-generation sequencing libraries. Forensic Science International: Genetics Supplement Series, 5, e276–e278. https://doi.org/10.1016/j.fsigss.2015.09.110

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