Single-cell 3′ RNA sequencing with 10x Genomics technology allows gene expression study at a single-cell level, profiling thousands to tens of thousands of cells in parallel. It captures cellular heterogeneity, identifies rare cell types, and discerns gene expression differences, enhancing understanding of cellular diversity. The process involves cell isolation into droplets, RNA capture and barcoding and cDNA synthesis, library preparation, pooling, sequencing, and bioinformatic analysis for single-cell transcriptomic profiling.
The Single Cell Gene Expression Flex assay by 10X Genomics, enables single-cell/nuclei RNA-seq libraries from formaldehyde-fixed cells and tissues, including FFPE blocks. Utilizing sequence-specific probe pairs for about 18,000 human and 19,000 mouse genes, this method accounts for RNA degradation. Sample multiplexing, facilitated by sample barcodes in probes, allows pooling of discrete cell populations, accommodating up to 16 samples and potentially analyzing 128,000 cells per GEM reaction.