Targeted Sequencing of 179 Genes Associated with Hereditary Retinal Dystrophies and 10 Candidate Genes Identifies Novel and Known Mutations in Patients with Various Retinal Diseases

Supplemental Method Targeted sequence capture and NGS 6 μg of genomic DNA was randomly fragmented with sizes mainly distributed between 250 and 300 bp. Adapters were ligated to both ends of the resulting fragments. DNA was then amplified by ligation-mediated PCR (LM-PCR), purified, and hybridized to the RDs189 array for enrichment, and non-hybridized fragments were then washed out. Both non-captured and captured LM-PCR products were subjected to quantitative PCR to estimate the magnitude of enrichment. Each captured library was sequenced by the Illumina Genome Analyser II platform following the manufacturer’s instructions (Illumina, San Diego, CA, USA). Raw image files were processed by the Illumina basecalling Software 1.7 for base calling with default parameters and the sequences of each individual were generated as 90 base paired-end reads.