Each sample DNA library is combined with QIAseq qPCR SYBR® Green Mastermix, aliquoted in technical triplicate across the QIAseq Library Quant Array, and analyzed with qPCR. Raw CT values are copied into the data analysis Excel template, and the software automatically calculates the library concentration or dilution factor.
The QIAseq Library Quant Array's high sensitivity and broad dynamic range enable the quantification of both the NGS-L1 and NGS-L2 libraries [A]. By contrast, Agilent's High Sensitivity DNA Kit (for use with the Agilent 2100 BioAnalyzer) quantified only the NGS-L1 library; with this kit, the NGS-L2 concentration was too low for quantification [B].
The serial dilutions of the DNA standard (5 sequential 10-fold dilutions) generate a standard curve. The sample library should fall within the standard curve.