Clinical Genomics Gothenburg is now offering 10x Chromium single cell RNA sequencing
Single cell RNA sequencing (scRNA-seq) has become the state‐of‐the‐art approach for unravelling the heterogeneity and complexity of RNA transcripts within individual cells at unprecedented resolution. This is performed by isolating single cells, capturing their transcripts, and generating sequencing libraries in which the transcripts are mapped to individual cells. The Clinical Genomics Gothenburg (CGG) NGS sequencing unit at the Core Facilities is now available for performing scRNA-seq using 10X Genomics Chromium controller. We are accepting new projects for scRNA-seq along with standard and complete Bioinformatic analysis.
The first protocol for performing single-cell RNA sequencing was published in 2009 by Tang et al. Since then, the scRNA-seq approaches has been used as a powerful method for molecular and cellular characterization of individual cells by exploring gene expression profiles at a single-cell resolution and has been successfully used in different fields such as cancer, developmental biology, immunology and neuroscience. Unlike the bulk RNA sequencing (RNA-seq) which identifies the presence and quantification of RNA based on averaged gene expression from all cells in a population, scRNA-seq provides unbiased, high-throughput, and high-resolution transcriptomic analysis of individual cells.
We perform scRNA-seq using the 10X genomics Chromium controller instrument, which uses advanced microfluidics to perform single cell partitioning and barcoding in a matter of minutes. Powered by Next GEM (Gel Beads in emulsion) technology, the Chromium Controller enables integrated analysis of single cells at massive scale.
Using 10x Genomics scRNA-seq technology, we can generate data to analyze transcriptomes on a cell-by-cell basis by using microfluidic partitioning method to capture single cells and prepare barcoded, next-generation sequencing cDNA libraries. Briefly, single cells, reverse transcription reagents, Gel Beads containing barcoded oligonucleotides, and oil are combined on a microfluidic chip to form reaction vesicles called GEMs. These GEMs are tagged with unique barcodes, and upon gel beads dissolving all the identically barcoded RT oligonucleotides enters the solution, where Reverse transcription of mRNA occurs. As a result, all cDNAs from a single cell contains the same barcode and these barcoded cells are used for library preparation followed by sequencing using NextSeq or NovaSeq. 500-10 000 individual cells can be profiled per sample resulting in 30,000 – 80,000 reads per cell.
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