Gene Expression Nebulas
A data portal of transcriptomic profiles analyzed by a unified pipeline across multiple species

Gene Expression Nebulas

A data portal of transcriptome profiles across multiple species

PRJNA322291: Characterisation of EZH2-deficient human embryonic stem cells [single cell RNA-seq]

Source: NCBI / GSE81673
Submission Date: May 20 2016
Release Date: Dec 06 2016
Update Date: May 15 2019

Summary: Here we analyse single cell transcriptome profiles of EZH2-deficient human embroynic stem cells

Overall Design: Single cell transcriptome (mRNA-Seq) from Ezh2-/- (Null) and EZH2+/+ (WT) human ESC

GEN Datasets:
GEND000207
Strategy:
Species:
Healthy Condition:
Cell Type:
Protocol
Growth Protocol: Human ESC (WA09; obtained from WiCell) were cultured on CF1 irradiated mouse embryonic fibroblasts (MEF) in Advanced DMEM containing 20% Knockout Serum Replacement (Life Technologies) supplemented with 2mM L-Glutamine (Life Technologies); 0.1mM B-Mercaptoethanol (Sigma-Aldrich); 1x Penicillin/Streptomycin (Life Technologies); 1x Non-Essential Amino Acids (Life Technologies) and 4ng/ml FGF2 (WT-MRC Cambridge Stem Cell Institute). Cells were passaged using 1mg/ml Collagenase IV (Life Technologies).
Treatment Protocol: -
Extract Protocol: SSEA4-positive live hESC were deposited by FACS directly into 96-well plates. Poly-A cDNA from single cells was obtained as described by Picelli et al.
Library Construction Protocol: Illumina Nextera DNA LT library preparation kit was used following the Fluidigm protocol described for cDNA generation using the Fluidigm C1 machine (available online). Spike-ins (ERCC RNA Spike-in Mix, Ambion) were added in a final dilution of 1:10E6.
Sequencing
Molecule Type: poly(A)+ RNA
Library Source:
Library Layout: PAIRED
Library Strand: -
Platform: ILLUMINA
Instrument Model: Illumina HiSeq 2500
Strand-Specific: Unspecific
Samples
Basic Information:
Sample Characteristic:
Biological Condition:
Experimental Variables:
Protocol:
Sequencing:
Assessing Quality:
Analysis:
Data Resource GEN Sample ID GEN Dataset ID Project ID BioProject ID Sample ID Sample Name BioSample ID Sample Accession Experiment Accession Release Date Submission Date Update Date Species Race Ethnicity Age Age Unit Gender Source Name Tissue Cell Type Cell Subtype Cell Line Disease Disease State Development Stage Mutation Phenotype Case Detail Control Detail Growth Protocol Treatment Protocol Extract Protocol Library Construction Protocol Molecule Type Library Layout Strand-Specific Library Strand Spike-In Strategy Platform Instrument Model Cell Number Reads Number Gbases AvgSpotLen1 AvgSpotLen2 Uniq Mapping Rate Multiple Mapping Rate Coverage Rate
Publications
Deletion of the Polycomb-Group Protein EZH2 Leads to Compromised Self-Renewal and Differentiation Defects in Human Embryonic Stem Cells.
Cell reports . 2016-12-01 [PMID: 27926872]