GSM4838252: corpus_P42; Mus musculus; RNA-Seq
The tissue pieces were digested in 2 ml GEXSCOPETM Tissue Dissociation Solution (Singleron Biotechnologies) at 37 ℃ for 15 min in a 15 ml centrifuge tube with continuous agitation. Following digestion, a 40-micron sterile strainer (Corning) was used to separate cells from cell debris and other impurities. To remove red blood cells, 2 mL GEXSCOPETM Red Blood Cell Lysis Buﬀer (Singleron) was added to the cell suspension and incubated at 25°C for 10 minutes. The mixture was then centrifuged at 1000 rpm for 5 min and cell pellet was re-suspended in PBS. Cells were counted with TC20 automated cell counter (Bio-Rad). Single cell suspension was loaded onto a microfluidic chip (part of Singleron GEXSCOPETM Single Cell RNAseq Kit, Singleron Biotechnologies) and single cell RNA-seq libraries were constructed according to manufacturer's instructions (Singleron Biotechnologies). The resulting single cell RNAseq libraries were sequenced on Illumina HiSeq X10 instrument with 150bp paired end reads.