GSM2484524: DGM-13206_Sample_76; Homo sapiens; RNA-Seq
Single cell capture, RNA extraction and cDNA preparation were performed on the Fluidigm C1 system following the manufacturer’s protocol. To enrich the cell capture of club cells and basal cells, and minimize the inclusion with larger differentiated ciliated and mucin-secreting cells, the single cell suspension (15 μl; 400~600 cells/μl) were loaded into the medium-sized (10 to 17 μm in diameter) microfluidic RNA-seq chip after priming (Fluidigm). The cells were captured using an Olympus-phase contrast microscope to examine the cell numbers in each capture site. Only single cells in the capture site were used. The Fluidigm C1 system was used for single cell RNA purification and cDNA synthesis by using the SMARTer Ultra Low RNA kit, and the single cell cDNAs were transferred to a 96 well-plate. The single cell library preparation and RNA sequencing were performed following a Fluidigm protocol. The single cell libraries were pooled and sequenced with paired end clustering and 101x2 cycles sequencing on the Illumina Hiseq 2500
GEO Accession
GSM2484524