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SRX6774371: GSM4048293: Knock_down3; Mus musculus; RNA-Seq
1 ILLUMINA (Illumina NovaSeq 6000) run: 21.1M spots, 5.9G bases, 1.8Gb downloads

Submitted by: NCBI (GEO)
Study: Gene expression changes following Knock-down of eRNA_06347 in post-stroke mouse cortex
show Abstracthide Abstract
RNA-sequencing was conducted to profile the transcriptome of the eRNA_06347 knockdown post-stroke mouse cortices ( knockdown achieved by intracerebroventricular injection of oligos against eRNA_06347). RNA was isolated from sham, Negative control oligo injected post-stroke cortices and eRNA_06347 knockown post-stroke cortices (three independent biological replicates per group),converted into cDNA libraries, and used for Illumina deep sequencing.The sequencing reads were aligned to the mouse reference genome GRCm38 using HISAT2. Stringtie was used to assemble the aligned sequences into transcript isoforms using mouse transcript annotations from Ensembl 81 as a guide, then merged to reconstruct a comprehensive transcriptome using perl scripts and gffcompare. After the ?nal transcriptome was generated, StringTie and Ballgown were used to estimate the expression levels of all the transcripts. StringTie was used to evaluate expression levels for mRNAs by calculating the FPKMs. The differentially expressed mRNAs were identified as log2 fold change >1 or log2 fold change <-1 and with statistical significance p<0.05 using Ballgown.Doing so, We found that 106 genes were significantly upregulated and 49 genes were significantly downregulated in the eRNA_06347 knockdown group as compared to the negative control. Overall design: Genome-wide transcriptomic profiles of eRNA_06347 knockdown post-stroke mouse cortices compared to sham cortices and negative control post-stroke mouse cortices
Sample: Knock_down3
SAMN12642321 • SRS5321471 • All experiments • All runs
Organism: Mus musculus
Library:
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Animals were euthanized and the infarcted ipsilateral cortical tissues were immediately harvested and snap-frozen in liquid nitrogen before storing in -80°C. Total RNA was then isolated from the tissues using the mirVana total RNA isolation kit, followed by Dnase I tratment. The RNA integrity was evaluated using Bioanalyzer. For Negative control and Knockdown samples Poly(A) RNA sequencing, the library was prepared following Illumina's truseq-stranded-mRNA sample preparation protocols. RNA integrity of total RNA sample received was checked using an Agilent Technologies 2100 Bioanalyzer. The polyA containing mRNA molecules were purified using poly-T oligo attached magnetic beads and using two rounds of purification. After purification the polyA RNA was fragmented using divalent cation containing buffer and elevated temperature. Quality control analysis and quantification of the DNA library were performed using Agilent Technologies 2100 Bioanalyzer High Sensitivity DNA Chip. Paired-end sequencing was performed on Illumina's NovaSeq 6000 sequencing system.For the Sham samples,previously generated data from our lab was used. Please refer to GEO accession number GSE112348 for the relevant details regarding sham samples.
Experiment attributes:
GEO Accession: GSM4048293
Links:
Runs: 1 run, 21.1M spots, 5.9G bases, 1.8Gb
Run# of Spots# of BasesSizePublished
SRR1003941821,120,2215.9G1.8Gb2020-11-12

ID:
8924038

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