Comparison of techniques for quantification of next-generation sequencing libraries

TY - GEN

T1 - Comparison of techniques for quantification of next-generation sequencing libraries

AU - Hussing, Christian

AU - Kampmann, Marie-Louise

AU - Mogensen, Helle Smidt

AU - Børsting, Claus

AU - Morling, Niels

PY - 2015/12/1

Y1 - 2015/12/1

N2 - 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.

AB - 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.

KW - Faculty of Health and Medical Sciences

KW - DNA quantification

KW - Next Generation Sequencing

KW - NanoDrop

KW - Qubit

KW - Fragment analyzer

KW - GX Touch

KW - Bioanalyzer

KW - TapeStation

KW - DNA quantification

KW - NanoDrop

KW - Qubit

KW - Next generation sequencing

KW - Fragment analyzer

KW - GX Touch

KW - Bioanalyzer

KW - TapeStation

U2 - 10.1016/j.fsigss.2015.09.110

DO - 10.1016/j.fsigss.2015.09.110

M3 - Conference article

SN - 1875-1768

VL - 5

SP - e276-e278

JO - Forensic Science International: Genetics. Supplement Series

JF - Forensic Science International: Genetics. Supplement Series

T2 - 26th Congress of the International Society of Forensic Genetics

Y2 - 31 August 2015 through 5 September 2015

ER -