QIAseq cfDNA All-in-One Kit (96)

A dedicated QIAGEN solution that combines highly efficient circulating cell-free DNA (cfDNA) extraction and next-generation sequencing (NGS) library preparation in a single workflow for any liquid biopsy sample. The kit pairs QIAamp Mini Column-based extraction with library construction reagents designed for dilute, low-input cfDNA, producing libraries suitable for analysis on Illumina and Ion Torrent sequencing instruments. Extraction supports up to 5 ml of plasma, yielding eluant from which 1–100 ng of cfDNA in up to 56.5 µl can be taken directly into library preparation without prior quantification.

The library preparation step combines end-polishing and adapter ligation in an all-in-one enzyme mix, with the ligation reaction itself completed in approximately 35 minutes and the full library prep taking less than one hour. Single-use barcoded sequencing adapters are pre-dispensed into a 96-well plate, enabling multiplex sequencing and reducing the risk of adapter cross-contamination. PCR-free libraries can be generated from as little as 10 ng of cfDNA input, with the workflow reliably reaching the 2 nM library concentration typically required for Illumina MiSeq or NextSeq 500 sequencing, and an optional high-fidelity library amplification step is provided for samples below 10 ng cfDNA.

High-efficiency adapter ligation chemistry delivers reproducible conversion of cfDNA to library and uniform exome coverage across varying input amounts, enabling sensitive variant detection down to 1% allelic frequency. Parts of the extraction protocol can be automated on QIAcube or QIAsymphony SP platforms with equivalent results, and the complete plasma-to-library protocol takes less than three hours.