Application note: A complete next-generation sequencing workflow for circulating cell-free DNA isolation and analysis

Circulating cell-free DNA (cfDNA) has been shown to have potential as a noninvasive substrate for the detection and monitoring of tumour cells. As circulating tumour DNA is often present at low frequencies within cfDNA, targeted sequencing is an optimal tool for mutation detection.

To support the advancement of cfDNA research, we demonstrate a complete workflow providing: isolation of cfDNA from plasma using the Applied Biosystems™ MagMAX™ Cell-Free DNA Isolation Kit either manually with a magnetic stand or automatically on the Thermo Scientific™ KingFisher™ Flex or KingFisher™ Duo Prime Magnetic Particle Processor, followed by molecular characterisation of isolated cfDNA using the multiplexing capabilities of Ion AmpliSeq™ technology and the Ion PGM™ System.

cfDNA isolated with the MagMAX Cell‑Free DNA Isolation Kit and amplified with the Ion AmpliSeq™ Cancer Hotspot Panel v2 demonstrates reproducible amplicon representation and variant detection across 50 genes of interest, covering 2,800 COSMIC mutations.

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