Single Cell Sequencing
For Single Cell Sequencing the facility is equipped with a Chromium Controller of 10x Genomics. With that technology, gene expression profiles of up to 10,000 single cells or nuclei can be generated using their proprietary droplet technology.
In the device, single cells are brought together with gel-bead loaded with barcoded oligonucleotides in a minute droplet called GEM (gel-bead in emulsion), in which cDNA is generated from the mRNAs of a single cell. All cDNA originating from a single droplet will have the same, unique barcode, which after sequencing, allows annotation of the reads to the single cell within this droplet and generate the single cell transcriptome. More detailed information on the proprietary 10xGenomics technology for single cell sequencing can be found on the company´s website (https://www.10xgenomics.com/products/single-cell-gene-expression).
Aside from living cells, also cells fixed with certain fixation protocols and isolated cell nuclei can be analysed using these workflows. We are happy to give advice to our user prior to their project and discuss options as well as sample requirements for the different workflows.
Aside from consultation and help with project planning, our service includes the library preparation from a used supplied single cell suspension including all process controls and sequencing using external sequencing service on an Illumina sequencer. The user is responsible for generation of the single cell suspension or single nuclei suspension that meet the sample requirements as discussed during consultation.