Amplicon sequencing of Pa in CF sputum
AmpliSeq libraries were constructed using the Ion AmpliSeqTM Library kit 2.0 and IonCodeTMbarcode set with the following modifications. SparQ magnetic beads (Quantabio) were used forpurification, and individual libraries were quantified using the Quant-iT PicoGreen dsDNAAssay Kit (ThermoFisher). Samples were mixed in equimolar proportions and the pooled library(200 pM) was loaded on an Ion Chef for template preparation using HiQ reagents. The P1 v3chips were sequenced using an Ion Proton sequencer (500 flows) with P1 HiQ sequencingreagents following manufacturers instructions