Single-Cell RNA-Seq Library Preparation

Single-cell suspensions (2 × 10⁵ cells/mL) with PBS (HyClone) were placed onto a microwell chip using the Singleron Matrix^® Single Cell Processing System. Reverse transcription of the mRNA captured by the Barcoding Beads is followed by PCR amplification of the cDNA generated from the Barcoding Bead collection. Sequencing adapters are then ligated to the fragmented cDNA. GEXSCOPE^® Single Cell RNA Library Kits (Singleron) were used to construct the scRNA-seq libraries (Dura et al., 2019 (link)). Illumina novaseq 6,000 was used to sequence 150 bp paired end reads from individual libraries diluted to 4 nM, pooled, and processed.