Ion torrent pgm hi q sequencing kit

Ion Torrent adapter-ligated DNA libraries were built according to the Ampliseq™ Library Preparation kit 2.0 96Lv (Thermo Fisher Scientific, Inc.) using 10 ng of input DNA from peripheral blood mononuclear cells (PBMCs), quantitated by Qubit 2.0 (Invitrogen; Thermo Fisher Scientific, Inc.) in two primer-pools with 118 and 115 amplicons, respectively. The polymerase chain reaction (PCR) products of the two pools were mixed together, with samples barcoded using Ion Xpress Barcodes 1–16 (Thermo Fisher Scientific, Inc.), followed by purification using AMPure XP beads (Beckman Coulter, Inc., Brea, CA, USA) and quantification by Ion Library TaqMan™ Quantitation kit (Thermo Fisher Scientific, Inc.). Prepared libraries were pooled in equal amount and diluted to 100 pM. Subsequently, 2 µl of pooled library were enriched using a Ion One-Touch Two (OT2) system with the Ion Personal Genome Machine™ (PGM) Hi-Q™ OT2 kit (Thermo Fisher Scientific, Inc.), followed by enrichment of Ion Sphere Particles on the Ion OneTouch Enrichment System apparatus (Thermo Fisher Scientific, Inc.). Enriched products were sequenced using the Ion Torrent PGM Hi-Q™ Sequencing kit using the Ion 316™ chip v2 (Thermo Fisher Scientific, Inc.).