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Gene Expression Profiling in Laser-Microdissected Bone Marrow Megakaryocytes

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Gene expression analysis of the megakaryocytic lineage requires isolation of megakaryocytes from their bone marrow microenvironment. Laser microdissection of megakaryocytes from diagnostic bone marrow biopsies allows analysis of standardised formalin-fixed samples that reflect the in situ grown status quo of a physiological or pathological condition. Taking into account that in neoplastic proliferation, e.g. myeloproliferative neoplasms, non-neoplastic haematopoietic clones proliferate in parallel, this morphology-based isolation enables selective analysis of the aberrant megakaryocytic population. Two different laser microdissection devices are presented, and the details of RNA extraction and sub-sequent real-time qPCR gene expression analysis of mRNA and microRNA are provided.
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