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Sample GSM4981580 Query DataSets for GSM4981580
Status Public on Oct 01, 2022
Title HB_TENT-5_3
Sample type SRA
 
Source name L4 worms
Organism Caenorhabditis elegans
Characteristics strain: tent-5(tm3504) I (NBRP)
genotype: tent-5(tm3504) mutant
treatment: grown at 25℃ on TSA plates seeded with E. coli HB101
tissue: whole worm
replicate: 3
Treatment protocol Synchronized wild-type and mutant worms were grown at 25°C on NGM plates seeded with E. coli HB101 until they reached the L4 stage. Worms were washed 3x with 50 mM NaCl and resuspended in 1 ml of TRI Reagent (Sigma-Aldrich)
Extracted molecule total RNA
Extraction protocol Total RNA from C. elegans and BMDM cells was isolated with TRI Reagent (Sigma-Aldrich) according to the manufacturer’s instructions. Before RNA-seq library preparation, total RNA samples were treated with TURBO DNase (Thermo Fisher Scientific) to remove DNA contamination. RNA was then purified by phenol/chloroform extraction and ethanol precipitation.
Ribosomal RNA was removed from samples using a Ribo-Zero rRNA Removal Kit (Human/Mouse/Rat, Epicentre). Strand-specific libraries were prepared using a dUTP protocol (KAPA Stranded RNA-Seq Library Preparation Kit), according to the manufacturer. Library quality was assessed using chip electrophoresis performed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Inc.).
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina NextSeq 500
 
Data processing Illumina bcl2fastq 2.20 software used for basecalling.
Sequence reads from C. elegans experiments were mapped to the WBCel235 genome build (ENSEMBL, release 94) using STAR v. 2.6.1b
Sequence reads from M. musculus were mapped to the GRCm38 mouse genome build (ENSEMBL, release 94) using STAR v. 2.7.6a
Counts for each transcript in C. elegans experiments were collected using featureCounts from Subread package (v. 1.6.3), with options options -Q 10 -p -B -C -s 2 -g gene_id -t exon and respective annotation gtf file (WBCel235; ENSEMBL; release 94)
Counts for each transcript in M. musculus experiments were collected using featureCounts from Subread package (v. 2.0.1), with options options -Q 10 -p -B -C -s 2 -g gene_id -t exon and respective annotation gtf file (Gencode, vM25)
Differential expression analysis was performed using DESeq2, with default settings
Genome_build: GRCm38
Genome_build: WBcel235
Supplementary_files_format_and_content: tab-delimited text files include DESeq2 normalized counts for each sample.
 
Submission date Dec 19, 2020
Last update date Oct 01, 2022
Contact name Paweł S Krawczyk
E-mail(s) pkrawczyk@iimcb.gov.pl
Organization name International Institute Of Molecular And Cell Biology In Warsaw
Lab Laboratory of RNA Biology
Street address 4 Ks. Trojdena Street
City Warsaw
ZIP/Postal code 02-109
Country Poland
 
Platform ID GPL19757
Series (1)
GSE163549 TENT5 cytoplasmic non-canonical poly(A) polymerases regulate the innate immune response in animals
Relations
BioSample SAMN17120520
SRA SRX9706759

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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