Illumina panel sequencing familial pancreatic cancer
The panel of 35 genes was designed using the Agilent SureDesign Software. 50ng of genomic DNA was fragmented and adaptors were added and the DNA library was purified and amplified. 750ng of each library were hybridized using SureSelectQXT capture library and recovered using streptavidin magnetics beads and a post-capture PCR amplification was carried out. Sequencing was performed on the MiSeq (Illumina) using paired-end protocol 2 x 150 bp and Micro V2 chemistry. On average, 5.1 million of pass filter reads were produced per sequencing run and 93.9% of reads had an average Phred score quality above Q30 and the read depth was greater than 200.