Advancing Autism Research in a Diverse Population: Exploring Autism Multiplex Data in Indian Families
Whole-Exome sequencing libraries were prepared using SureSelectXT Human All Exon (V5). Biotinylated oligonucleotide capture probes, also called as baits, that were designed for the human exons were provided with the kit and used to enrich the region of interest (whole exome) by hybridization. The workflow involved shearing of DNA, repairing ends, adenylation of 3 ends, followed by adapter ligation. At each step, the products were purified. The adapter sequences were added onto the ends of DNA fragments to generate paired-end libraries. The resulting adaptor ligated libraries were purified, qualified, and hybridized with an exome-specific biotinylated capture library. After hybridization, the targeted molecules were captured on streptavidin beads. The resulting enriched DNA libraries were multiplexed by adding index tags by amplification, followed by purification. Indexed captured library DNA was assessed to check the quality and quantity of the captured libraries. Finally, indexed captured library DNA was sequenced on Illumina HiSeq series to generate 2X150bp sequence reads at 80 - 100X on target sequencing depth.