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Protocols: Bone marrow mononuclear cells were stained with conjugated monoclonal antibodies for HSPC markers as described above except for CD34 in order to minimize staining time and prevent RNA degradation (CD34 staining requires 90 minutes (Okamoto et al., 2007). Single cells were sorted into 96-well plates with the lysis buffer and stored frozen at -80. cDNA amplification and sequencing were performed as per Smart-Seq2 protocol (Picelli et al., 2013). Multiplexed sequencing libraries were prepared from cDNA using the Illumina Nextera XT protocol.
BioProject SRA
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