Single-cell RNA Sequencing from Microwell Chip

Single-cell suspensions (2×10⁵ cells/mL) with PBS (HyClone) were loaded onto microwell chip using the Singleron Matrix^® Single Cell Processing System. Barcoding Beads are subsequently collected from the microwell chip, followed by reverse transcription of the mRNA captured by the Barcoding Beads and to obtain cDNA, and PCR amplification. The amplified cDNA is then fragmented and ligated with sequencing adapters. The scRNA-seq libraries were constructed according to the protocol of the GEXSCOPE^® Single Cell RNA Library Kits (Singleron) (23 (link)). Individual libraries were diluted to 4 nM, pooled, and sequenced on Illumina novaseq 6000 with 150 bp paired end reads.