Sureselect xt target

In the initial pilot phase, commercially available SureSelect XT Human-All-Exome RNA baits (Agilent Technologies Inc.) were used following the instruction from the manufacturer.
The retrospective cohort was interrogated, using a novel panel of 266 genes. These genes were uploaded into the Agilent SureSelect E-array software to design 56,008 probes (3× tiling density), using moderately stringent masking. The probes covered exons (± 10 nt.), 3′ UTRs and 5′ UTRs of the 266 candidate genes under investigation. Total genomic region spanned 1.357 Mbp. The sequences of all 56,008 baits are listed in the Supplementary File 1. Biotinylated RNA baits were synthesised by Agilent Inc. for the SureSelect XT Target Enrichment system.

We performed whole-exome sequencing using the SureSelect XT Target enrichment system and the SureSelect Human All Exon V6 bait (Agilent). We obtained 2 × 150-bp paired-end sequencing data using a HiSeq2500 next-generation sequencing platform to cover the exome with 100× coverage, and variant calling was performed essentially in the same way as whole-genome sequencing.