PND120_pro (CRX1243833)
The scRNA-seq libraries were generated using the 10x Genomics Chromium Controller Instrument and Chromium Single Cell 3' V3.1 Reagent Kits (10x Genomics, Pleasanton, CA). The process was as follows: 1) Cells were concentrated to approximately 1000 cells/uL and loaded into each channel to generate single-cell Gel Bead-In-Emulsions (GEMs). 2) After the reverse transcription (RT) step, the GEMs were broken and barcoded-cDNA was purified and amplified. 3) The amplified barcoded cDNA was fragmented, subjected to A-tailed, and ligated with adaptors followed by index PCR amplification. 4) The final libraries were quantified using the Qubit High Sensitivity DNA assay (Thermo Fisher Scientific), and the size distribution of the libraries were determined using a High Sensitivity DNA chip on a Bioanalyzer 2200 (Agilent). 5) All libraries were sequenced by Illumina sequencer (Illumina, San Diego, CA) on a 150 bp paired-end run.