Whole Exome seq of cell line: pedi DIPG: Sample DIPG13_sgMSH6_DMSO
Library construction was performed as described in Fisher et al., with the following modifications: initial genomic DNA input into shearing was reduced from 3µg to 10-100ng in 50µL of solution. For adapter ligation, Illumina paired end adapters were replaced with palindromic forked adapters, purchased from Integrated DNA Technologies, with unique dual-indexed molecular barcode sequences to facilitate downstream pooling. With the exception of the palindromic forked adapters, the reagents used for end repair, A-base addition, adapter ligation, and library enrichment PCR were purchased from KAPA Biosciences in 96-reaction kits. In addition, during the post-enrichment SPRI cleanup, elution volume was reduced to 30µL to maximize library concentration, and a vortexing step was added to maximize the amount of template eluted.