GSM5570892: Sham 3; Mus musculus; RNA-Seq
Islets were isolated from pancreata using Collagenase digestion and Histopaque gradient separation, hand-picked, dissociated using Trypsin, FACS-sorted to single cells and seeded separately in 384 well-plates with lysis buffer. Single cells are lysed, and the 3' ends of mRNAs are annealed to UMIs containing a T7 promoter. The mRNA is reverse-transcribed to cDNA and amplified by IVT. The RNA strands are fragmented and annealed to sequencing adapters, followed by RT to generate barcoded cDNA libraries that are ready for sequencing. single-cell MARSseq