GSM3135439: 7527AA_MSCB-098; Mus musculus; RNA-Seq
Short-term cultured pancreatic islet cells were isolated from a healthy pancreas donor. The culturing process favor the enrichment of pancreatic islet cells while large eliminating endothelial and immune cell types. Primary bladder epithelial cells were isolated by FACS (CD45-, CD140a-, CD31-, Ter119-) from a healthy mouse. To construct single cell libraries from polyA-tailed RNA, we applied massively parallel single-cell RNA sequencing (MARS-Seq; Jaitin et al., Science 2014). This methodology generates paired-end reads where read 1 contains the transcript sequence and read 2 the cell and UMI barcodes (bases 1 to 6 and 7 to 14, respectively). Briefly, single cells were FACS-sorted into 384-well plates, containing lysis buffer and reverse-transcription (RT) primers. The RT primers contained the single cell barcodes and unique molecular identifiers (UMIs) for subsequent de-multiplexing and correction for amplification biases, respectively.