RNA sequencing (RNAseq) provides a more complete genetic picture than DNA sequencing. It is one of the most frequently and widely used tools in Omic oncology. RNAseq includes the classic bulk RNAseq, the popular single cell RNA sequencing (scRNAseq) and the emerging spatial RNA sequencing (spRNAseq). Bulk RNAseq investigates global RNA biology in a tissue or cell population, while scRNAseq refines it to a single cell level, and spRNAseq has the additional ability to dissect cell-cell communications spatially. The driving force for the evolution of these technologies was specific oncological questions and technological limitations. Thus, each of these tools has its unique features and suitable applications in precision oncology. It is desirable for cancer researchers to recognize their respective strengths, weaknesses, and optimal applications.
Workflow of 10X Genomics Single Cell Sequencing
The procedures include seven key steps. The library construction from step 1 to 5) takes 2 days. Novaseq 100 cycle kits are normally used for sequencing. Sequencing sets up as read1:28 cycles for cell barcode & UMI, i7 index: 8 cycles for sample index, read 2: 91 cycles for insert. Cell Ranger, Loupe Browser and other commercial or publicly available software are used for data analysis.