2025-09-10
ENA first public
ERC000011
ENA-CHECKLIST
Leibniz Institute for Natural Product Research and Infection Biology Hans Knöll Institute (Leibniz-HKI)
INSDC center name
public
INSDC status
ERS24422308
SRA accession
E-MTAB-15110:Sample 7
Submitter Id
ArrayExpress
broker name
dendritic cell
cell type
not collected
collection date
human
common name
Protocols: Monocyte-derived dendritic cells (moDCs) were generated from CD14+ monocytes isolated from peripheral blood mononuclear cell as previously described.  Seelbinder B, Wallstabe J, Marischen L, Weiss E, Wurster S, Page L, Löffler C, Bussemer L, Schmitt A-L, Wolf T, Linde J, Cicin-Sain L, Becker J, Kalinke U, Vogel J, Panagiotou G, Einsele H, Westermann AJ, Schäuble S, Loeffler J. 2020. Triple RNA-Seq Reveals Synergy in a Human Virus-Fungus Co-infection Model. Cell Reports 33:108389.  Guinan J, Lopez BS. 2020. Generating Bovine Monocyte-Derived Dendritic Cells for Experimental and Clinical Applications Using Commercially Available Serum-Free Medium. Frontiers in Immunology Volume 11 - 2020. Total RNA from the samples was isolated according to the manufacturer’s instructions. An on-column DNase digestion step was included in the RNA isolation protocol. RNA extraction kit: RNeasy Mini/Micro Kit (Qiagen) Elution volume: 40 µl Library preparation was performed with the NEBNext® Poly(A) mRNA Magnetic Isolation Module and NEBNext® UltraTM II Directional mRNA kit (New England Biolabs) according to the manufacturer’s instructions (2). The protocol starts with a poly(A) RNA selection step using Oligo dT Beads d(T). The mRNA was fragmented and samples were then introduced  into a reverse transcription to generate first strand cDNA. After the second strand cDNA  synthesis, 3’-ends were adenylated and sequencing adapters were ligated. Those comprise  sequencing primer- and flow cell-binding sites, as well as indices for multiplexed sequencing  of pooled libraries. Adapter-ligated fragments were amplified during a 15-cycle PCR reaction. Universal Human Reference RNA (UHR) was simultaneously processed as internal positive  control (PosC) but discarded after successful library validation.
description
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
blood
organism part
Homo sapiens
scientific_name
cIL10_Afu
stimulus
Sample 7
title