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Design: scRNA-seq of index-sorted myeloid and lymphoid progenitor cells and differentiated pDCs
Submitted by: Institute for Immunology, LMU Munich
Study: scRNA-seq of index-sorted myeloid and lymphoid progenitor cells and differentiated pDCs
show Abstracthide Abstract
Nine different cell types (common dendritic progenitor (CDP), pre-conventional dendritic cell (pre-cDC), common dendritic progenitor (CDPr, according to Rodrigues et al., but this population was flawed in sorting purity), Flt3+ CD11c+ Siglec-H+ CCR9-low B220-low progenitor cell (lo-lo), Flt3 + CD11c+ Siglec-H+ CCR9-low B220-high progenitor cell (lo-hi), plasmacytoid dendritic cell (pDC), Ly6D+ lymphoid progenitor (SP), Ly6D+ Siglec-H+ lymphoid progenitor (DP) and common lymphoid progenitor (CLP)) were sorted to allow for analysis of their transcriptomic relation and/or similarity. The pDC_precursor_scvelo.h5ad file is a processed file ready for direct downstream analysis with scvelo.
Library:
Name: AM_SC4_1_s
Instrument: Illumina HiSeq 1500
Strategy: OTHER
Source: TRANSCRIPTOMIC SINGLE CELL
Selection: RT-PCR
Layout: SINGLE
Construction protocol: Lineage-positive cells were depleted from the bone marrow (BM) cell suspension by magnetic cell sorting using APC-Cyanine7 antibodies against CD3, CD19, Ly6G, NK1.1, Ter119 and anti-Cy7 beads (Miltenyi Biotech) according to the manufacturer's protocol followed by incubation with viability dyes and antibodies against cell surface markers diluted in FACS buffer (PBS, 2% FCS, 2 mM EDTA) for 20 minutes on ice in the dark. After washing with FACS buffer cells were resuspended in FACS buffer and sorted on a BD FACSAria Fusion as single cells into 96-well plates containing 5µl mcSCRB-seq lysis buffer (5M Guanidine HCl, 1% β-ME, 0.2% Phusion HF buffer 5x) per well. mRNA was extracted according to the mcSCRB-seq protocol (lysis buffer: 5M Guanidine HCl, 1% β-ME, 0.2% Phusion HF buffer 5x) ref: doi: 10.1038/s41467-018-05347-6 Libraries were generated according to the mcSCRB-seq protocol. ref: doi: 10.1038/s41467-018-05347-6
Experiment attributes:
Experimental Factor: cell type: pre-cDC, lo-lo, lo-hi, pDC, CLP, CDPr
No Runs

ID:
21949014

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