GSM8581896: Ctrl; Mus musculus; RNA-Seq
Cells were washed with 1× PBS detached with 0.25% Trypsin incubated for 3 minutes. Then cells were washed twice with 1× PBS with 1% FBS and resuspended in cold 1× PBS with 1% FBS. Cells were filtered through a 40 µm cell strainer and centrifuged at 500 g for 5 min at 4°C. Pellets were resuspended in cell resuspension buffer at 1000 cells per μl for library preparation. The DNBelab C Series Single-Cell Library Prep Set (MGI) was used as previously described for single-cell RNA sequencing library preparation. In brief, single-cell suspensions were converted to barcoded scRNA-seq libraries through steps including droplet generation, emulsion breakage, mRNA captured bead collection, reverse transcription, cDNA amplification and purification. Indexed sequencing libraries were constructed according to the manufacturer's protocol. The sequencing libraries were quantified by Qubit ssDNA Assay Kit (Thermo Fisher Scientific). Libraries were sequenced on the DNBSEQ platform at the China National GeneBank (Shenzhen, China) according to the manufacturer's protocol.