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
Strategy: |
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Species: |
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Healthy Condition: |
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Cell Type: |
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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. |
Molecule Type: | poly(A)+ RNA |
Library Source: | |
Library Layout: | PAIRED |
Library Strand: | - |
Platform: | ILLUMINA |
Instrument Model: | Illumina HiSeq 2500 |
Strand-Specific: | Unspecific |
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 |
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