Dataset View [GSE73727]

TitleSingle-cell transcriptomics reveals unique features of human pancreatic islet cell subtypes
ArticleKubicek S,Bock C,Berishvili E,Barbieux C,Spittler A,Penz T,Farlik M,Klughammer J,Li J.Single-cell transcriptomes reveal characteristic features of human pancreatic islet cell types.EMBO reports.2016 Feb
Bio ProjectBioProject: http://www.ncbi.nlm.nih.gov/bioproject/PRJNA297774
SraSRA: http://www.ncbi.nlm.nih.gov/sra?term=SRP064464
Overall DesginTranscriptional dissection of human pancreatic islets of one donor using single-cell RNA-seq
SummaryWe report the single-cell RNA-seq based identification of 6 known human islet cell types (alpha cells, beta cells, delta cells, pp cells, acinar cells and duct cells) based on the expression of known marker genes. We further assess cell type specific gene expression and suggest novel marker genes for several cell types.
Experimental ProtocolTo disassociate islets into single cells, islets were incubated in Accutase (Life technology) in 37 °C for 20 min, neutralized by CMRL medium. Purification of single cells was performed by flow cytometry cell sorting on a Moflo AstriosEQ (Beckman Coulter, Miami); cDNA synthesis and enrichment was performed following the Smart-seq2 protocol as described (Picelli et al., 2014).
Data processingTrimming of adapter sequences was performed with trimmomatic (v 0.32) minimum read length after trimming: 25; Alignment of the trimmed reads to the human transcriptome (hg19 GRCh37 ftp://ftp.ensembl.org/pub/release-74/fasta/homo_sapiens/cdna/Homo_sapiens.GRCh37.74.cdna.all.fa.gz) was performed with bowtie1 (v 1.1); Calculation of normalized transcript-wise expression estimates (RPKM values) was performed based on the transcriptome alignments using the R (v 3.1.2) package BitSeq (v 1.10.0). Function: getExpression() with uniform=FALSE; Genome_build: hg19; Supplementary_files_format_and_content: Tab-delimited text files include transcript-wise RPKM values or read counts for each sample
Public OnPublic on Jan 05 2016

Cell Groups

Differential Expression Gene List

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