The development of new sequencing platforms, technologies, and bioinformatics tools in the past decade fostered key discoveries in human genomics. Among the most recent sequencing technologies, nanopore sequencing (NS) has caught the interest of researchers for its intriguing potential and flexibility. This up-to-date review highlights the recent application of NS in the hematology field, focusing on progress and challenges of the technological approaches employed for the identification of pathologic alterations. The molecular and analytic pipelines developed for the analysis of the whole-genome, target regions, and transcriptomics provide a proof of evidence of the unparalleled amount of information that could be retrieved by an innovative approach based on long-read sequencing. Since the longer a read is, the more overlap it has with other reads, third generation sequencing technology facilitates the assembly of genomic regions and could resolve complex and wide structural variants or regions enriched for repetitive elements.Nanopore sequencing (NS) provides significant advantages for whole-genome studies and, thanks to the ability to span huge regions, it could be exploited to screen specific genes of pathologic relevance affected by unknown rearrangements (i.e., F8, BCR-ABL) or composed of repetitive elements and high GC content (i.e., CEBPA, FLT3).Owing to the nature of NS technology, the ability to direct sequence-untouched nucleic acids minimizes the bias associated with PCR amplification and enables the simultaneous detection of nucleotide sequence and the epigenetic changes of the template.The rapid workflow required to prepare NS libraries, along with the possibility to set real-time data analysis, allows retrieval of information of clinical utility at whole-genome and target region level in a unique reaction in 24/48 h.