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Single Cell RNA-Seq

Single Cell RNA-Seq

Single cell RNA-Seq (scRNA Seq) is a tool that enables simple and robust access to the transcriptomes of thousands of single cells – giving unprecedented insight into tissues at the level of individual cells. This offers vital information and data and is key to understanding many diseases and immunity.

This method enables analysis of millions of single cells in a high-throughput manner. This is in contrast to traditional techniques, that either allowed analysis of a few genes in thousands of individual cells (e.g., in situ hybridization), or the expression profile of thousands of genes, but only on a tissue homogenate.

Single cell RNA-Seq enables new types of analyses, for instance:

  • Understanding tumour heterogeneity and clonal evolution – including studying lineage analysis, cancer stem cells, and drug resistant and metastatic clones.
  • Understanding complex tissues (e.g. neural tissues).
  • High resolution identification of cells types and markers, and understanding differentiation pathways in developmental and systems biology.

To meet the demand for simple and robust, high throughput, single cell transcriptomics, reliable droplet-based protocols for single cell RNA-Seq were published in 2015 (Macosko E., et al. “Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets.” Cell 161:1202). These allow thousands of cells to be captured individually in droplets, together with lysis reagents and a barcoded bead, all within in a few minutes. Inside a droplet, an individual cell is lysed, and its mRNA is captured on uniquely barcoded primers attached to the bead; the beads are recovered from the emulsion and subject to bulk reverse transcription, library preparation, and sequencing. Each cell’s transcriptome can be identified by clustering by barcode.

Dolomite Bio has developed systems to execute the Drop-seq (described in Macosko et al., 2015) and other, related protocols, leading to rapid adoption worldwide.  Dolomite Bio’s RNA-Seq system allows cells to be efficiently captured into droplets at high speed.

High precision, inert, glass chips are connected to pressure-based pumps to create a reliable system under computer control, with real time imaging of cell encapsulation and droplet formation to give confidence in system performance.

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