Accurate quantification of library molecules is essential for the efficient use of NGS platforms; however, current spectrophotometric methods only measure total nucleic acid concentrations. The QIAseq Library Quant System, by contrast, uses real-time PCR to quantify only DNA molecules with adaptors at both ends, which are the only amplifiable molecules during emulsion PCR (Ion Torrent platform) or bridge PCR (Illumina platform), and therefore provides highly accurate quantification of amplifiable library molecules. The high sensitivity of real-time PCR allows quantification of libraries at very low concentrations, even below the detection threshold of conventional spectrophotometric methods.
The QIAseq Library Quant System is available in both a tube and an array format, and is optimized with QIAseq qPCR SYBR
® Green Mastermixes to provide superior sensitivity and a wide dynamic range. Both formats provide a DNA standard specific to the Illumina or Ion Torrent platform. The array format provides this standard in five predispensed, sequential 10-fold dilutions mixed with a PCR primer assay in triplicate, ensuring that your sample library will fall within the detection range of the array (see figure
Principle of QIAseq Library Quant System).