Targeted NGS of Pateint 16
A custom AmpliSeq panel with 336 primer pairs (avg. amplicon length: 352 bp) was designed using Illuminas DesignStudio, covering 98.45% of the target region. DNA (10 ng) was quantified using the Qubit dsDNA HS Assay and amplified with 5x AmpliSeq HiFi mix. Post-amplification, FuPa digestion, pooling, CD index ligation, and AMPure XP cleanup were performed. Libraries were quantified via Qubit and assessed on a TapeStation (avg. size: 400550 bp). Final libraries (~10 pM) were sequenced using paired-end reads on the Illumina MiSeq platform.