Comparison of techniques for quantification of next-generation sequencing libraries

被引：10

作者：

Hussing, C. ^{[1
]}

Kampmann, M. L. ^{[1
]}

Mogensen, H. S. ^{[1
]}

Borsting, C. ^{[1
]}

Morling, N. ^{[1
]}

机构：

[1] Univ Copenhagen, Fac Hlth & Med Sci, Dept Forens Med, Sect Forens Genet, Copenhagen, Denmark

来源：

FORENSIC SCIENCE INTERNATIONAL GENETICS SUPPLEMENT SERIES | 2015年 / 5卷

关键词：

DNA quantification; Next-generation sequencing libraries; NanoDrop; Qubit; (R); Fragment Analyzer (TM); GX Touch; Bioanalyzer; TapeStation;

D O I：

10.1016/j.fsigss.2015.09.110

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

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 (TM) and Illumina (R) 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 Qubit1 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. (C) 2015 Elsevier Ireland Ltd. All rights reserved.

引用

页码：E276 / E278

页数：3