NextSeq 500 paired end sequencing; GSM1501785: CC10- B4+ single cells and K5-Cre+ single cells; Mus musculus; RNA-Seq

CC10- B4+ single cells and K5-Cre+ single cells

We used microfluidic single cell RNA-seq on adult isolated CC10-CreERT2 (negative) integrin beta4(pos) cells lung epithelial cells in order to determine the transcriptional profile of this putative progenitor population. Overall design: CC10-CreERT2 / tdTomato (negative) integrin beta4(pos) cells were isolated by FACS, as were Krt5-CreERT2 / tdTomato (positive) cells.  These cells were pooled and loaded onto the Fluidigm C1 device.

High throughput quantitative whole transcriptome analysis of CC10- B4+ and Krt5-CreERT2 - labeled distal lung cells

Submitted by Gene Expression Omnibus on 26-NOV-2014

Cells were loaded into the Fluidigm C1 microfluidic platform, where single cells were captured. Lysis of single cells, reverse-transcription of mRNA into cDNA as well as preamplification of cDNA occured within the microfluidic device using reagents provided by Fluidigm as well as the SMARTer Ultra Low RNA kit for Illumina (Clontech). External RNA spike-in transcripts (ERCC spike-in Mix, Ambion) were added to all single cell lysis reactions. As a bulk control, cDNA was prepared from approximately 200 cells in a PCR tube using the same protocol and reagents as used for the single cells. In addition, one "no cell control" was prepared using reagents without any cell input in a PCR tube. Libraries were prepared using Illumina Nextera XT kit per illumina's protocols.

High throughput quantitative whole transcriptome analysis of CC10- B4+ and Krt5-CreERT2 - labeled distal lung cells

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