Illumina HiSeq 2500 sequencing; GSM1818992: TRA00028923_ls_14; Mus musculus; RNA-Seq

GEO accession GSM1818992 is currently private and is scheduled to be released on Jul 14, 2015.

Single cell transcriptomic analysis of wildtype and AireKO thymic epithelial cells Overall design: Single cells were sorted by FACS for single cell RNAseq library preparation

Single cell transcriptomic analysis of thymic epithelial cells

Submitted by Gene Expression Omnibus on 17-JUL-2015

Thymi were removed and digested in collagenase/dispase to single cell suspensions. Medullary thymic epithelial cells were sorted by FACS as CD45-Ly51-MHCII+GFP+. Single cells were sorted into water in wells of 96well plate and lysed by freeze-thaw. mRNA was reverse transcribed using adapters containing oligo dT and ArrayScript (Ambion), second strand synthesis was performed using Second strand synthesis module (NEBNext #E6111L), and linear amplification was performed by in vitro transcription using MEGAshortscript T7 transcription kit (Ambion AM1354). Illumina RNA adapters were ligated onto transcripts using T4 RNA Ligase 2, truncated (Enzymatics L6070L) and processed according to standard Illumina protocols.

Single cell transcriptomic analysis of thymic epithelial cells

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