GSM7431637: brain, PRVABC-59; Mus musculus; RNA-Seq
Briefly, single cells were suspended in PBS containing 0.04% BSA. Approximately 10,000 cells were added to each channel and approximately 6,000 target cells were recovered. Captured cells were lysed and the released RNA was barcoded through reverse transcription in individual GEMs. Reverse transcription was conducted on a S1000TM Touch Thermal Cycler (BioRad) at 53 °C for 45 min, followed by 85°C for 5 min and a hold at 4°C. Complementary DNA was generated and amplified, after which, quality was assessed using an Agilent 2100 (performed by Bestopcell, Beijing, China) According to the manufacturer's introduction, scRNA-seq libraries were constructed using a Chromium Next GEM Single Cell 3ʹ Kit v3.1 (10X Genomics, 1000268) The libraries were sequenced using an Illumina Novaseq 6000 sequencer with a paired-end 150-bp (PE150) reading strategy (performed by Bestopcell, Beijing, China)