GSM9081751: Cre-Vhlh-KO; Mus musculus; RNA-Seq
The sorted cells from Cre-Vhlh-KO and Cre-WT were centrifuged and resuspended in 1X PBS with 0.04% BSA. Cell viability and concentration were assessed using trypan blue staining and an automated cell counter ensuring >90% viability. Single-cell suspensions were loaded into the 10x Genomics Chromium Single Cell A Chip according to the manufacturer's protocol (Chromium Single Cell 3' Reagent Kits v3 User Guide, CG000183). Single-cell RNA libraries were prepared using the Chromium Single Cell 3' Library & Gel Bead Kit v3 (10x Genomics, PN-1000075), following the manufacturer's protocol. Briefly, single cells were encapsulated into droplets along with barcoded gel beads using the Chromium Controller. Following reverse transcription and cDNA amplification, libraries were constructed by fragmentation, end repair, A-tailing, adapter ligation, and PCR amplification. The resulting libraries were quantified and quality-checked using a Bioanalyzer (Agilent Technologies) and sequenced on an Illumina NovaSeq 6000 platform with paired-end 150 bp reads.