RNA-Sequencing and targeted exome sequencing of venetoclax resistant human leukemia from spleens and bone-marrow of patient-derived xenograft models

For RNAseq, mRNA was extracted from spleen and bone marrow of venetoclax resistant and matched parental PDX leukemia cells using RNeasy kit (Qiagen). Non-stranded RNA-seq libraries were generated using the True-SeqRNA exome library prep kit (Illumina) on a Sciclone platform (Perkin Elmer). mRNA underwent fragmentation, cDNA synthesis, and next-generation library synthesis via exome capture and PCR amplification. Libraries were sequenced on a Next-Seq instrument (Illumina) using a paired-end protocol. For targeted exome sequencing using rapid heme panel assay, genomic DNA was extracted from spleen and bone marrow of venetoclax resistant and matched parental PDX leukemia cells. DNA underwent customized hybrid-capture target enrichment sequencing followed by UMI correction. Variants were called using Vardict.