| Summary: | Associated Persons Michael Murrey (Creator); Eloise Greenland (Creator)James Steer (Creator) 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_M10_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 ...
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