Methods for characterisation of individual cells using a wide range of solutions.
Bulk RNA transcriptomic methods have become very popular and give access to basic gene expression (GE) data on eg. whole tissues. However, any tissue is going to be composed of multiple cell types, and cells with a somewhat different agenda. To really dissect gene expression signals at the cellular level, one may need to look at gene expression at the single cell level.
A number of different methods for single cell expression analysis are available, each with their own advantages and disadvantages. The data you get out may be from the full transcript or the 5’ / 3’ ends (eg. 10X), the platform may be optimised for multi-omics info from each cell (eg. 10X with cell surface marker info or ATAC data) or for getting you data from the largest possible number of cells (eg. the Parse 1M kit). You may prefer fixing cells (eg. 10X Flex & Parse) or bringing us freshly isolated cells/nuclei (10X 5’ / 3’ kits). Finally, the sequencing depth and the optimal scale to plan for - these will depend on what you are after.
NGI currently offers single-cell RNA sequencing using the droplet-based 10X Genomics and the split-plate Parse platforms, but we are also continually evaluating new methods and protocols. On the single-cell DNA sequencing side, we have tested kits from Bioscryb and Qiagen, contact us if you want to know more about that!
Multiple rounds of barcoding of fixed and permeabilised cells.
mRNA illumina RNA-Seq single-cell parse combinatorial transcriptomics RNACombining short & long reads at the single cell level.
ont chromium 10x Genomics full transcript library preparation single cell long-read illuminaProfiling of chromatin accessibility at the single cell level.
chromium chromatin 10x Genomics ATAC library preparation epigenetics single cell illumina ATAC-seq dnaProfiling of 3´gene expression and chromatin accessibility in the same cell.
RNA mRNA illumina ATAC-seq RNA-Seq ATAC chromium library preparation 10x Genomics transcriptomics single cellProfiling of gene expression levels at single-cell resolution.
chromium 10x Genomics library preparation transcriptomics single cell RNA mRNA illumina RNA-SeqSingle-cell profiling of gene expression levels on fixed cells.
transcriptomics single cell RNA mRNA illumina RNA-Seq chromium 10x Genomics fixed cells library preparation