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E-MTAB-8882

ABSTRACT: RNA sequencing of primary isolated unstimulated murine splenic cDC from specific pathogen free (SPF), Germ Free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice.

PROVIDER: E-MTAB-8882 | ExpressionAtlas | 2021-01-19

REPOSITORIES: ExpressionAtlas

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			Action	DRS
	E-MTAB-8882-analysis-methods.tsv	Tabular
	E-MTAB-8882-analytics.tsv	Tabular
	E-MTAB-8882-atlasExperimentSummary.Rdata	Rdata
	E-MTAB-8882-configuration.xml	Xml
	E-MTAB-8882-g3_g1-mvaPlot.png	Other

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RNA sequencing of primary isolated unstimulated murine splenic cDC from specific pathogen free (SPF), Germ Free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice.

Project description:The transcriptome of primary isolated unstimulated murine splenic cDC from specific pathogen free (SPF) was compared to cDC from Germ Free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice. Cells were isolated and sorted by flow cytometry to high purity. Total RNA was isolated using the RNeasy plus micro kit (Qiagen). Barcoded mRNA seq libraries were prepared from 100 ng of total RNA (RIN>8) using a NEBnext poly(A) mRNA Magnetic Isolation Module and NEBnext UltraII lib prep kit for Illumina (New England Biosciences) according to the manual. Single-end RNA sequencing (59nt) was performed on a HiSeq 2500 (Illumina). Barcoded RNA-seq libraries were onboard clustered using HiSeq® Rapid SR Cluster Kit v2 using 8pM and 59bps were sequenced on the Illumina HiSeq 2500 using HiSeq Rapid SBS Kit v2 (59 Cycle). The raw output data of the HiSeq was preprocessed according to the Illumina standard protocol.

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RNA sequencing of primary isolated unstimulated murine splenic DC1 and DC2 from specific pathogen free (SPF), Germ free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice

Project description:The transcriptomes of primary isolated unstimulated murine splenic cDC1 and cDC2 from specific pathogen free (SPF) were compared to cDC1 and cDC2 from Germ Free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice. Total RNA was prepared from sort-purified cDC1 and cDC2 (for each sample 3 individual mice were pooled) using Trizol reagent (Invitrogen) in combination with the miRNeasy Micro Kit (Qiagen) according to the manufacturer’s instructions. For poly-A-dependent cDNA synthesis and a first amplification step 10 ng of total RNA was used as input in the Smart-Seq v4 mRNA Ultra Low Input RNA Kit (Clontech) and processed according to the manufacturer’s instructions. 1 ng of the purified cDNA was used for tagmentation and library completion with the Nextera XT library preparation kit (Illumina). In the following, 2x75nt paired-end sequencing was performed on a NextSeq 500.

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E-MTAB-8883

Project description:RNA sequencing of primary isolated unstimulated murine splenic DC1 and DC2 from specific pathogen free (SPF), Germ free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice

2024-03-04 | E-MTAB-8883 | ExpressionAtlas

RNA sequencing of primary isolated unstimulated murine splenic cDC from specific pathogen free (SPF), Germ Free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice.

Project description:RNA sequencing of primary isolated unstimulated murine splenic cDC from specific pathogen free (SPF), Germ Free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice.

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