title: Microbiome sample from the blood of a surgical patient
design: Suppression adapters were generated from a long and short oligonucleotide (Sigma-Aldrich, Steinheim). Both oligonucleotides were incubated with 10 L 10x T4 ligase mix (New England Biolabs, Frankfurt) and 50 L nuclease-free water for 3 min at 95 C and cooled down overnight at RT. SUPSETS workflow starts with ligating suppression adapters to blunt-end DNA fragments (prepared cfDNA), followed by elongation in the presence of a panel of Fusion primers. For clinical samples, a defined volume of 15.92 L blunt-end cfDNA was used. After AMPure XP beads purification, second PCR amplification was performed using US-FW and Ad-rev primers (both Sigma-Aldrich, Steinheim). Purified amplification products were applied to Nanopore sequencing. For MinION Flow Cell experiments, the Native Barcoding Kit 24 V14 (Oxford Nanopore Technologies, ONT, London) was used according to manufacturers advice and sequenced for 72 husing MinKnow. The basecalling was done using guppy in high accuracy mode.