GSM9036106: Sample1; Mus musculus; RNA-Seq
Single-cell selection were performed using the SMARTer ICELL8 Single-Cell System (Takara Bio) per manufacturer's instructions. In brief, single cell suspension was filtered with a 100-µm nylon cell strainer. Afterwards, cells were stained with a mixture of Hoechst 33342 and propidium iodide and counted. Then, after dilution, a cell suspension of 20,000 cells/ml was submitted to the MultiSample NanoDispenser for single cell preparation. Cells were dispensed into an ICELL8 Chip, and imaged on the Imaging Station, from where live, single cells were selected by Hoechst-positive and Propidium Iodide-negative staining. Reverse transcription and amplification were performed on a Chip Cycler (Bio-Rad), followed by cDNA purification and size-selection with Agencourt AMPure XP beads. The libraries were generated with the Nextera XT Library Preparation Kit (Illumina, FC-131-1024) following the manufacturer's instructions, and then sequenced on the NovaSeq 6000 sequencer