To understand a system, you need to understand its components. If you look at complex biological systems, like the immune system or a growing tumor, understanding the function and behavior of the different cells in these systems is a big challenge. With single-cell RNA sequencing, scientist now have a great and powerful tool to face this challenge. In contrast to traditional bulk RNA sequencing, where gene expression levels are averaged across cells, single-cell RNA sequencing allows you to look at the gene expression of thousands of individual cells. Single-cell RNA sequencing is the perfect technique for all applications, where understanding cellular diversity is essential, like studying tumor heterogeneity, cellular differentiation, or therapy resistance.
At CeGaT, we provide high throughput single-cell RNA sequencing using the state-of-the-art Chromium system from 10x Genomics in combination with the latest NGS technology. The sample processing workflow includes three main steps: cell barcoding & library construction, sequencing, and data analysis. During cell barcoding, up to 10.000 single cells are encapsulated into tiny micro-droplets together with gel beads. These gel beads contain oligonucleotides with cell-specific barcodes and unique molecular identifiers. By labelling all mRNA molecules of a single cell with these two barcode sequences, their cellular origin is recorded, and each mRNA molecule can be uniquely quantified.