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Sample GSM4558309 Query DataSets for GSM4558309
Status Public on Oct 22, 2020
Title NIC1_S6: NPCs grown inside the Nichoid
Sample type SRA
 
Source name Sub Ventricular Zone
Organism Mus musculus
Characteristics cell type: Neural Precursor Stem Cells (NPCs)
treatment: nichioid
Treatment protocol NIC samples were harvested, collected by centrifugation (123xg for 10 minutes), mechanically dissociated and counted with trypan blue reagent (Sigma Aldrich). 10 000 cells were seeded at the centre of the Nichoid in a single drop of 35 μL NSCs medium. The multi-well was kept in the incubator for 1 hour to allow cells to enter the 3D niches and then 465μL of NSCs medium were added.
Growth protocol NSCs were maintained in culture in Neurobasal Medium (GIBCOTM, Life Technologies) containing 2% B-27 supplement (Life Technologies), 2% L-Glutamine (Euroclone), 1% penicillin and streptomycin (Euroclone), b-FGF (human recombinant, 20 ng/mL) and hEGF (human recombinant, 20 ng/mL).
Extracted molecule total RNA
Extraction protocol Neural Precursors Cells (NPCs) expressing green fluorescent protein (GFP) were isolated 6 h post-mortem from adult C57BL/6-Tg(UBC-GFP)30Scha/J mice weighing 25–30 g (Charles River)
Sequencing libraries were prepared by the Illumina TruSeq Stranded Total RNA kit (Illumina) using 200-ng total RNA (Illumina). Qualities of sequencing libraries were assessed with 4200 TapeStation with the DNA1000 reagent kit. RNA processing has been carried out using Illumina NextSeq 500 Sequencing.
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina NextSeq 500
 
Data processing FastQ files were generated via llumina bcl2fastq2 Version 2.17.1.14
Gene and transcript intensities were computed using STAR/RSEM software using Gencode Release m24 (GRCm38) as a reference, using the “-strandness forward” option. Differential expression analysis for mRNA was performed using R package DESeq2
Genome_build: M24
Supplementary_files_format_and_content: RSEM output: transcript_id, gene_id, length, effective_length, expected_count, TPM, FPKM
 
Submission date May 18, 2020
Last update date Oct 22, 2020
Contact name Stella Gagliardi
E-mail(s) stella.gagliardi@mondino.it
Organization name IRCCS Mondino Foundation
Department GPG center
Street address Via Mondino, 2
City Pavia
State/province Pavia
ZIP/Postal code 27100
Country Italy
 
Platform ID GPL19057
Series (1)
GSE150767 Dissecting the effect of a 3D micro-scaffold on the transcriptome of Neural Stem Cells with computational genomics
Relations
BioSample SAMN14945271
SRA SRX8358238

Supplementary file Size Download File type/resource
GSM4558309_NIC1_S6.gtf_annotated_genes.results.txt.gz 1.4 Mb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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