C3_R3_48h_IL4_Mature macrophage_mouse3

Sample type: SRA

Source name: bone marrow derived
Organism: Mus musculus
Characteristics
strain: C57BL/6
Sex: male
age: 8 to 10 weeks
Growth protocol: After 1 day in 0.6ng/ml CSF1 in alpha+ MEM /15% FCS, non-adherent cells were incubated for two days in a fresh dish containing 12ng/ml CSF1 in alpha+ MEM /10% FCS and then for 7 days in 120ng/ml CSF1 in alpha+ MEM /10% FCS. Cells were incubated for a further two days in fresh alpha+ MEM /10% FCS containing 120ng/ml CSF1 and 20ng/ml IL4.
Extracted molecule: total RNA
Extraction protocol: mRNA was harvested using RNeasy kit( QIAGEN) with DNase treatment on column. 1 ug of total RNA was used for the construction of sequencing libraries.
RNA libraries were prepared for sequencing using standard Ion Torrent protocols

Library strategy: RNA-Seq
Library source: transcriptomic
Library selection: cDNA
Instrument model: Ion Torrent S5

Description: IL4_48h_M26_BMM
Wt_vs_IL4_allprobes_reads.txt
Wt_vs_IL4_allprobes_log2_RPM.txt
Data processing: Torrent Suite Software 5.10 used for basecalling and sequenced reads were trimmed for adaptor sequence, and masked for low-complexity or low-quality sequence- returning a fastq file (raw data)
Reads were then mapped to the GRCm38.p6 genome using the open source Hisat2-2.0.5 aligner.
The Hisat2 generated BAM files were uploaded into SeqMonk (version 1.42) with minimum mapping quality set to 60
The edgeR platform, within SequeMonk, was uesed to generate lists of differential gene expression from the raw reads as is required in analysis of negative binomial distributions
Tab-delimited text files of all genes and differentially expressed genes (at p<0.05, p<0.01 and p<0.001) showing raw reads or log2 RPM were output (processed files)
Ampliseq
Torrent Suite Software 5.10 used for basecalling and sequenced reads were trimmed for adaptor sequence, and masked for low-complexity or low-quality sequence. Returning a fastq file (raw data) of reads associated with each of the 16000 barcoded primer pairs.
Reads were then mapped to the GRCm38.p6 genome using the open source Hisat2-2.0.5 aligner.
The Hisat2 generated BAM files were uploaded into SeqMonk (version 1.42) with minimum mapping quality set to 60
The edgeR platform, within SequeMonk, was uesed to generate lists of differential gene expression from the raw reads as is required in analysis of negative binomial distributions
Tab-delimited text files of all genes and differentially expressed genes (at p<0.05, p<0.01 and p<0.001) showing raw reads or log2 RPM were output (processed files)
Genome_build: Genome Reference Consortium mouse genome (GRCm39.p6)
Supplementary_files_format_and_content: tab-delimited text files include reads or log2 RPM for each sample showing all genes or differential expression between conditions.