QIAseq Library Quant Assay kit

Part of the QIAseq Library Quant System, the QIAGEN QIAseq Library Quant Assay Kit provides qPCR-enabled quantification of next-generation sequencing (NGS) libraries. The kit uses real-time PCR to specifically quantify DNA molecules carrying sequencing adaptors at both ends, supporting optimal cluster density for Illumina sequencing or template-to-bead ratio for Ion Torrent/Proton sequencing.

Two workflow formats are available. In the array format, the sample library is prepared using two tenfold dilutions, then mixed with QIAseq qPCR SYBR Green Mastermix and aliquoted in technical triplicate across the array plate. In the tube format, five sequential tenfold dilutions of DNA Standard are prepared alongside two tenfold dilutions of the sample library, then combined with the provided PCR assay and QIAseq qPCR SYBR Green Mastermix. After PCR, CT values are exported to a provided data analysis Excel file for automatic calculation of library concentration (Illumina) or template dilution factor (Ion Torrent/Proton).

A manufacturer comparison showed the system capable of quantifying libraries at concentrations below the detection limit of Agilent's High Sensitivity DNA Kit on the 2100 BioAnalyzer.