Ion Torrent library preparation and sequencing
Ten nanograms of genomic DNA extracted from peripheral blood were used for library preparation. DNA was amplified using the two gene panel primer pools. PCR pools for each sample were combined and indexed using the Ion Xpress Barcode Adapters kit. The amplified libraries were quantified using the qPCR through the Ion Library TaqMan™ Quantitation Kit.
All samples were diluted at 10 pM, then amplicon libraries were pooled for emulsion PCR and chip loading using the Ion Chef system, according to manufacturer’s instructions. The final pool of samples was sequenced with Ion GeneStudio S5 system using Ion 510 or Ion 520 Chips.