Qiagen, 333314, QIAseq Library Quant Assay Kit

1 tube of Primer Mix (500 µl), 1 bottle of Dilution Buffer (30 ml), 1 tube DNA Standard (100 µl), and 5 tubes GeneRead qPCR SYBR Green (1.35 ml) for sample library quantification prior to NGS

Features

- Ready-to-use, predispensed, sequentially diluted DNA standard (array format)
- Automatic calculation of library concentration
- Simple protocol, high sensitivity, and wide dynamic range
- Compatibility with Illumina and Ion Torrent/Proton NGS platforms
- Compatibility with most qPCR instruments

Product Details

The QIAseq Library Quant System provides a simple, out-of-the-box solution to quantify NGS libraries, enabling consistent results with every NGS run. The QIAseq Library Quant System uses real-time PCR to specifically quantify DNA molecules with sequencing adaptors at both ends, ensuring optimal cluster density or template-to-bead ratio.

Please note : GeneRead Library Quant Array (product number 180611) and GeneRead Library Quant Kit (product number 180612) will be phased out by the end of 2016. Please use QIAseq Library Quant Assay Kit (product number 333314) instead.

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Performance

The sensitivity and dynamic range of the QIAseq Library Quant System were compared against Agilent's High Sensitivity DNA Kit, using two NGS libraries of differing concentrations. The first library was quantifiable by both systems, but the low concentration of the second library was below the detection limit of Agilent's 2100 BioAnalyzer. By contrast, the QIAseq Library Quant Array was able to quantify both libraries (see figure QIAseq Library Quant System enables quantification of libraries with concentrations below the detection limit of conventional methods ).

Procedure

The array format workflow (see figure QIAseq Library Quant Array workflow ) begins with two tenfold dilutions of the sample library, to ensure that its concentration falls within the range of the serially diluted standards. The samples are mixed with QIAseq qPCR SYBR Green Mastermix and aliquoted in technical triplicate across the array plate. PCR is performed, and raw C T values are exported to the provided data analysis Excel file for automatic calculation of library concentration (Illumina platform) or template dilution factor (Ion Torrent/Proton platform).

The tube format workflow is similar to the array format workflow, with a few small differences. The procedure begins with preparation of five sequential tenfold dilutions of DNA Standard and two tenfold dilutions of the sample library. This ensures that the concentration of the library will fall within the range of the standard dilutions. Next, the diluted DNA standards and sample libraries are mixed with the provided PCR assay and the appropriate QIAseq qPCR SYBR Green Mastermix. PCR is performed, and C T values are exported to the provided data analysis Excel file for automatic calculation of library concentration (Illumina platform) or template dilution factor (Ion Torrent/Proton platform).