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SRX515332: GSM1366197: CRD_4; Mus musculus; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2000) run: 51.8M spots, 10.5G bases, 6.4Gb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: Deep sequencing reveals cell-type specific patterns of single cell transcriptome variation
show Abstracthide Abstract
We present high quality deep read-depth single cell RNA sequencing for 91 cells from five mouse tissues and 18 cells from two rat tissues, along with 30 control samples of bulk RNA diluted to single-cell levels. We find that transcriptomes differ globally across tissues with regard to the number of genes expressed, the average expression patterns, and within cell-type variation patterns. We develop methods to filter genes for reliable quantification and to calibrate biological variation. All cell types include genes with high variability in expression, in a tissue-specific manner. We also find evidence that single cell variability of neuronal genes in mice is correlated with that in rats consistent with the hypothesis that levels of variation may be conserved. Overall design: From an initial 143 cells we identified 107 high quality samples with deep genic coverage, including 13 brown adipocytes, 19 cardiomyocytes, 19 cortical pyramidal neurons and 18 hippocampal pyramidal neurons from embryonic mouse, 8 cortical pyramidal neurons and 8 hippocampal pyramidal neurons from embryonic rat, and 22 serotonergic neurons from adult mouse. While mouse data is collected from three strains of mice at different ages, each cell type dataset is internally consistent. There are no technical replicates of single-cell samples. We additionally prepared 30 control samples, amplifications of bulk total cardiomyocyte RNA diluted to single cell quantities.
Sample: CRD_4
SAMN02725923 • SRS591221 • All experiments • All runs
Organism: Mus musculus
Library:
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Ventricular cardiomyocytes were isolated from E14.5 transgenic mice expressing the green S/G2/M fluorescent ubiquitination-based cell cycle indicator, “Fucci” (A. Sakaue-Sawano et al., Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. Cell 132, 487–498 (2008)), using a modified protocol of the neomyts cardiomyocyte isolation kit (Cellutron Life Technologies). The dissociated cell suspensions were resuspended in 500μl of PBS supplemented with 5% FBS solution and sorted by flow cytometryon a FACSAria instrument operating at low pressure (20 psi) using a 100μm nozzle. Samples were sorted individually into 96-well plates containing RT buffer for linear in vitro-transcription T7-based antisense RNA (aRNA) amplification. Collected samples were individually amplified using three rounds of a linear in vitro-transcription-based method. Samples were prepared for multiplexed paired-end sequencing using the Illumina mRNA-Seq system according to manufacturer instructions.
Experiment attributes:
GEO Accession: GSM1366197
Links:
External link:
Runs: 1 run, 51.8M spots, 10.5G bases, 6.4Gb
Run# of Spots# of BasesSizePublished
SRR123304251,841,96810.5G6.4Gb2015-05-21

ID:
714979

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