GSM4565562: HY_E14; Mus musculus; RNA-Seq
Hypothalamus was derived from Rax-CreERT2::Ai14 mice induced at E9.5, microdissected at both embryonic and postnatal days (E11.5, E14.5, P0 and P7) and dissociated into single-cell suspension. Live tdTomato+ cells were purified by fluorescent-activated cell sorting for single-cell transcriptomic sequencing(10x Genomics). Single-cell RNA-seq libraries were constructed  according to the manufacturer's protocol accompanying single cell 3 'Library and Gel Bead Kit V3 (10xGenomics, 1000075). Cell suspensions (300-600 living cells per microliter determined by Count Star) were loaded on a Chromium Single Cell Controller (10x Genomics) to generate single-cell gel beads in the emulsion. Captured cells were lysed and the released RNA were barcoded through reverse transcription in individual GEMs (Zheng et al., 2017). Using a S1000TM Touch Thermal Cycler (Bio Rad) to reverse transcribe, the GEMs were programed at 53°C for 45 min, followed 85°C for 5 min, and hold at 4°C. The cDNA was generated and then amplified, and the quality was assessed using the Agilent 4200 (performed by CapitalBio, Beijing). Finally sequencing was performed on the Illumina Novaseq6000 sequencer following the manufacturer's introduction. scRNA-Seq
GEO Accession
GSM4565562