title: SASP156
library_construction_protocol: The whole genome sequencing libraries were prepared using Twist Library Preparation Enzymatic Fragmentation kit. In brief, 50ng of DNA was enzymatically fragmented by targeting 250bp fragments size using a fragmentation reagent and the DNA fragments undergo end repair where in the mix converts the overhangs resulting from fragmentation into blunt ends. The 3' to 5' exonuclease activity of end repair mix removes the 3' overhangs and polymerase activity fills in the 5' overhangs. To the blunt ended fragments, adenylation was performed by adding single 'A' nucleotide to the 3' ends. To the adenylated fragments, Illumina universal adapters were ligated and purified with AMPure XP beads (Catalog: A63881, Beckman Coulter). Furthermore, the DNA was amplified by 8 cycles of PCR with KAPA HiFi HotStart Ready Mix and Twist UDI Primers. The amplified product was then purified using 0.9X AMPure XP beads (Catalog: A63881, Beckman Coulter) and the final DNA library was eluted in 15μl of 0.1X TE buffer. The above libraries were quantified using the Qubit dsDNA HS kit and exome enrichment was done using Twist Fast Hybridization Target Enrichment Protocol by pooling 8 samples and 6 samples (making total of 4 pools; (8*7=56;4*1=4), starting from 1500 ng of total gDNA library mass. The final enriched libraries were eluted in 15μl of 0.1X TE buffer.