Summary: Find the casual relationship between gene expression network and cellular phenotype at single cell resolution. We collected donated human pre-implatation embryos; and the embryonic stem cells derived from them; isolate individual cells; prepared single cell cDNAs; and sequenced them by HiSeq2000. Then we analyzed the expression of known RefSeq genes.
Overall Design: Development and applications of single-cell transcriptome analysis. Nat Meth 8; S6-S11.
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Growth Protocol: | Donated human preplantation embryos were dissociated into individual blastmeres and were separately amplied to set up single cell cDNA libraries. |
Treatment Protocol: | - |
Extract Protocol: | Amplied single cell cDNAs were further amplied for another ten cycles of PCR. Then it was sonicated into 200~500bp fragments ,then the standard TruSeq DNA library preparation kit was used following the manufacturer suggestion protocol |
Library Construction Protocol: | Amplied single cell cDNAs were further amplied for another ten cycles of PCR. Then it was sonicated into 200~500bp fragments ,then the standard TruSeq DNA library preparation kit was used following the manufacturer suggestion protocol |
Molecule Type: | poly(A)+ RNA |
Library Source: | |
Library Layout: | SINGLE |
Library Strand: | - |
Platform: | ILLUMINA |
Instrument Model: | Illumina HiSeq 2000 |
Strand-Specific: | Unspecific |