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.