Cytogenetic analyses in the AMLCG trials were performed centrally, and risk groups were defined according to the 2010 UK Medical Research Council (MRC) and the European LeukemiaNet (ELN) 2017 genetic risk classification (ELN2017). Patients were characterized for NPM1 and CEBPA mutations, FLT3 internal tandem duplications (FLT3-ITD), and KMT2A (formerly MLL) partial tandem duplications (KMT2A-PTD) using standard methods described recently.16 (link) Targeted amplicon sequencing of 68 recurrently mutated genes as published recently was used for genetic characterization in training set 1 and the validation set.17 (link) RNAseq libraries were prepared using the Sense mRNA Seq Library Prep Kit V2 (Lexogen, n=238) and the TruSeq RNA Library Preparation V2 Kit (Illumina, n=12). Between 500–1000 ng total RNA [RNA integrity number (RIN) >7] were used as input material. All sequencing was paired end and performed using polyadenylated-selected and, in case of the Lexogen libraries, stranded RNA sequencing. Processing details and sequencing metrics are provided in the Online Supplementary Appendix. Samples were sequenced on a HiSeq 1500 instrument (Illumina) as 100 bp reads to a targeted depth of 20 million mappable paired reads per sample according to the “Standards, Guidelines and Best Practices for RNA-Seq v.1.0 (June 2011)”18 recommendations of the ENCODE Consortium. Samples were aligned with STAR 2.4.019 (link) to the human hg19 reference genome and analyzed by DESeq2.20 (link) Details regarding the workflow are provided in the Online Supplementary Appendix.