title: Synthetic sample H. sapiens + P.aeroginosa
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 (digested genomic DNA), followed by elongation in the presence of a panel of Fusion primers. As input, either 4,000 and 6,000 genome equivalents of adapter ligated, digested pathogen or human DNA, respectively, 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. Samples for technical validation were sequenced for a total of 24 h using MinKnow. The basecalling was done using guppy in high accuracy mode.