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Using the Oxford Nanopore cDNA platform, we generated nearly 2 billion long reads (median 25.8 million per sample) from 71 human samples, including 48 acute myeloid leukemia or myelodysplastic syndrome samples, 25 of which had splicing-factor gene mutations (in\n                  <jats:italic>SRSF2, U2AF1<\/jats:italic>\n                  , or\n                  <jats:italic>SF3B1<\/jats:italic>\n                  ). An additional 23 samples were from sorted hematopoietic cell populations from healthy individuals. Using this dataset, we created a transcript assembly containing 174,162 novel isoforms that are not described in the reference transcriptome. Deep-scale proteomic validation confirmed that many of these transcripts are translated into protein. We also identified isoforms enriched in spliceosome-mutant samples and found proteomic evidence of frequent nonsense-mediated decay regulation of novel transcripts. This dataset is a valuable community resource, enabling detection of new transcripts in short-read data sets. An interactive portal to explore splicing patterns in these data is available at\n                  <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"https:\/\/leylab.org\/isoforms\/\">https:\/\/leylab.org\/isoforms\/<\/jats:ext-link>\n                  .\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Key Points<\/jats:title>\n                  <jats:p>Long-read sequencing enables the detection of many novel transcripts in AML, including many from splice-factor-mutant patient samples<\/jats:p>\n                  <jats:p>This expanded transcriptome is a valuable community resource and can be used to improve analyses of short-read RNAseq data<\/jats:p>\n                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