Analytical and clinical validation of an amplicon-based next generation sequencing assay for ultrasensitive detection of circulating tumor DNA
Fig 1
Overview of the LiquidHALLMARK assay workflow.
(A) Stabilized whole blood is collected using custom collection kits and cell-free DNA (cfDNA) isolated from plasma. Up to 20 ng of cfDNA is used for library preparation. (B) Targeted genomic regions are tagged with oligonucleotide barcodes and enriched via high efficiency amplicon-based (AmpliMark) library preparation before being sequenced to a median coverage of >8000X. UMI, unique molecular identifier. (C) Sequencing data is bioinformatically deconvoluted, with molecular barcodes enabling error suppression rates above 100X. (D) Somatic variants and other biomarkers are quantitative reported, with comprehensive interpretation encompassing drug actionability and clinical trial eligibility.