Genomics Platform
The Genomics Platform is able to provide the production of single-cell-RNA-seq & single-nuclei-RNA-seq library preparations using two different approaches, providing researchers the ability to choose the method most suited to their experiments.
As sample preparation is key to a successful scRNA-seq/snRNA-seq experiment, we are able to advise users on experimental planning and sample preparation to provide the best possible results. Libraries will be returned along with a QC report, and if required, advice on sequencing strategy and supplier.
As well as single-cell/nuclei library preparations the Genomics Platform hosts a user-operated GridION Long-Read sequencer and a Promega RSC instrument providing automated DNA/RNA extractions from a wide variety of sample types.
scRNAseq/snRNAseq
Droplet-based
10X Chromium – We can provide single-cell/single-nuclei assays available via the 10X Chromium including: including Single Cell Gene Expression, Single Cell Immune Profiling, Single Cell ATAC and single-cell multi-omics. Please contact for details and costings.
10X Genomics Chromium workflow: (A) The Chromium instrument and cartridge. (B) Schematic of the GEMs used to capture sequences of interest. GEMs are gel beads with capture primers consisting of a Illumina R1 read primer, 10X Barcode (cell ID), UMI (transcript ID), and a polyd(T) or capture sequence. (C) General 10X Genomics workflow. Cells/nuclei are partitioned with GEMS, cells/nuclei are lysed and RNA is barcoded, the resulting Barcoded DNA fragments are then made into sequence-ready libraries.
Plate-based split-seq
Parse Whole Transcriptome – A new and innovative plate-based approach to scRNA-seq & snRNA-seq with a flexible layout allowing researchers to profile from 10,000 to 1,000,000 cells, far higher than droplet-based methods. Instead of loading cells/nuclei directly into a microfluidic device, cells/nuclei are fixed using a proprietary kit. The fixed cells/nuclei can be used directly or stored at -80°C for up to 6 months before library preparation, which is ideal for time-courses or when sampling is infrequent. Please contact for details and costings.
Parse Biosciences WT methodology: (A) A single-cell/nuclei suspension is fixed, after which fixed suspensions can be stored before proceeding to barcoding. Barcoding and library preparation is followed by sequencing and data analysis. (B) Barcoding; fixed cells/nuclei undergo several rounds of split-pool barcoding to uniquely label transcripts according to cell of origin. Cells are then lysed, and barcoded transcripts undergo library preparation, sequencing, and data analysis. (C) With each round of split-pool barcoding, the total number of barcode combinations grows exponentially. (D) Single cell transcriptomes are constructed by grouping transcripts containing the same four barcode combinations.
Long-read sequencing
ONT GridION long-read sequencer – The GridION is able to sequence five libraries via five MinION flowcells at once. Featuring real-time base-calling and analysis the GridION is able to perform Adaptive Sampling to read only the genes of interest, greatly enriching the final dataset with the sequences users are interested in.
Lab automation
Promega RSC instrument – The Promega RSC instrument is a simple to operate automated DNA/RNA extraction device, which can process up to 48 samples in a single short run from a wide variety of sources.
Contact
Professor Dan Macqueen
Genomics Facility Academic Lead
Contact details
Related links
https://www.parsebiosciences.com/
https://nanoporetech.com/products/gridion
https://www.promega.co.uk/products/lab-automation/automated-dna-rna-extraction-purification-maxwell/