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  WI-38 human lung fibroblast cells

Overall DesignA microfluidic device that pairs sequence-barcoded mRNA capture beads with individual cells was used to barcode cDNA from individual cells which was then pre-amplified by in vitro transcription in a pool and converted into an Illumina RNA-Seq library. Libraries were generated from ~600 individual cells in parallel and extensive analysis was done on 396 cells from the U87 and MCF10a cell lines and from ~500 individual cells with extensive analysis on 247 cells from the U87 and WI-38 cell lines. Sequencing was done on the 3'-end of the transcript molecules. The first read contains cell-identifying barcodes that were present on the capture bead and the second read contains a unique molecular identifier (UMI) barcode, a lane-identifying barcode, and then the sequence of the transcript.
SummarySingle cell transcriptomics has emerged as a powerful approach to dissecting phenotypic heterogeneity in complex, unsynchronized cellular populations. However, many important biological questions demand quantitative analysis of large numbers of individual cells. Hence, new tools are urgently needed for efficient, inexpensive, and parallel manipulation of RNA from individual cells. We report a simple microfluidic platform for trapping single cell lysates in sealed, picoliter microwells capable of “printing” RNA on glass or capturing RNA on polymer beads. To demonstrate the utility of our system for single cell transcriptomics, we developed a highly scalable technology for genome-wide, single cell RNA-Seq. The current implementation of our device is pipette-operated, profiles hundreds of individual cells in parallel with library preparation costs of ~$0.10-$0.20/cell, and includes five lanes for simultaneous experiments. We anticipate that this system will ultimately serve as a general platform for large-scale single cell transcriptomics, compatible with both imaging and sequencing readouts.type = Expression profiling by high throughput sequencing
Dataset viewGSE66357
PMID26047807

Samples in WI-38 human lung fibroblast cells

Displaying 41-45 of 45 results.
SeriesSampleInstrumentOrganismTitleCell Source
GSE66357GSM1683323Illumina NextSeq 500Homo sapiensPS041-2_R2_1_829WI-38 human lung fibroblast cells
GSE66357GSM1683324Illumina NextSeq 500Homo sapiensPS041-2_R2_1_927WI-38 human lung fibroblast cells
GSE66357GSM1683325Illumina NextSeq 500Homo sapiensPS041-2_R2_1_928WI-38 human lung fibroblast cells
GSE66357GSM1683326Illumina NextSeq 500Homo sapiensPS041-2_R2_1_931WI-38 human lung fibroblast cells
GSE66357GSM1683327Illumina NextSeq 500Homo sapiensPS041-2_R2_1_96WI-38 human lung fibroblast cells

Gene rank in WI-38 human lung fibroblast cells

Displaying 21-30 of 18975 results.
Rank orderGene SymbolEnsembl ID
21RPL18AENSG00000105640
22TIMP1ENSG00000102265
23ACTG1ENSG00000184009
24RPL23ENSG00000125691
25RPS12ENSG00000112306
26TMSB10ENSG00000034510
27MT-ND3ENSG00000198840
28TPT1ENSG00000133112
29FTH1ENSG00000167996
30COL1A2ENSG00000164692