Single Cell Sequencing

Single Cell Service Overview

Offering both microfluidics-based and combinatorial-barcoding single cell methods

Transcriptomics of individual cells can allow researchers to gain insights into cellular heterogeneity, identify rare cell types, and uncover new biomarkers for diseases.

The SAGC Offers an End-to-End Single Cell Service - From Project Consultation Through to Bioinformatics Analysis.

Complete toolkit from 10X Genomics, including Chromium X and Chromium Controller
Certified Service Provider for Parse Biosciences
Enabling MGI Single Cell (DNBelab) - Only Genomics Facility in Australia with both the MGI DNBSEQ-T7 and DNBSEQ-G400

Read Our Single Cell Guide For Help Planning Your Experiment

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SAGC offers the complete range of 10X Chromium Applications

SAGC has a Chromium Controller and Chromium X Series from 10X Genomics to enable multiomic analysis through droplet GEM-based single cell partitioning and sequencing.

Applications available:

Single cell RNA expression (GEX)
Single cell Immune Profiling (5' with TCR/BCR options)
Single cell ATAC
Single cell ATAC + GEX
Single cell RNA FLEX (FFPE compatible)
Single cell CNV
Cell surface marker tagging can be incorporated into some of these techniques and quantified during sequencing.

Chromium GEM-X is NOW AVAILABLE

Updates to the reagent chemistry and microfluidics for both the 3’ Single Cell Gene Expression and the 5’ Immune Profiling solutions.

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Single Cell/Nuclei RNA Sequencing Services

Parse Biosciences Certified Service Provider

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The SAGC is a Certified Service Provider for Parse Biosciences and their Evercode™ technology. Evercode is a plate-based technology (requires no specialised equipment) and provides a highly scalable and flexible method to analyse individual cells or nuclei.

The assay is available in four formats that accommodate experimental designs of any size and scale.

	Mini	WT	Mega	Penta
Max Number of Cells	10K	100K	1M	5M
Max Number of Samples	12	48	96 or 384	384

Technology Overview

Evercode split-pool combinatorial barcoding from Parse Biosciences profiles single cell transcriptomes without the need for microfluidic instruments.

The high-level process is very similar for users familiar with other single-cell RNA sequencing technologies.

Create a single cell or nuclei suspension
Fix the samples
Prepare libraries
Sequence the libraries
Process the data via a pipeline before further analysis

How Combinatorial Barcoding Works

Parse Evercode technology differs from droplet-based single cell technology by using the cell or nuclei as the reaction vessel. There is no microfluidics instrument required.

Fixed and permeabilized cells/nuclei are loaded into a 96 well Round 1 Plate that is preloaded with well-specific barcodes. The RNA is reverse transcribed in situ with barcoded oligo dT and random hexamer primers.
The cells/nuclei are pooled and loaded into a second 96 well plate. An adaptor with a well-specific barcode is ligated to the first barcode. This is repeated to add a third barcode.
Pooled cells/nuclei are divided across several sublibraries and lysed. A fourth sublibrary-specific barcode is added during sequencing library preparation.
After sequencing, each transcript is assigned to a single cell/nuclei based on a unique combination of the four barcodes.

How Fixation Works

A key aspect of the Parse Biosciences Evercode technology is sample fixation, which is applicable to a wide range of animal cells/nuclei. Cells/nuclei are fixed in about 65-70 minutes, and fixed samples are stable for up to six months. This means you can work on a time course study, share samples between laboratories, or accumulate samples before analyzing them all together.

Assays

Evercode^TM Single Cell Technology: Evercode allows researchers to conduct scalable single cell experiments with no microfluidics instrument required. The assay is available in four formats that accommodate experimental designs of any size and scale.

Whole Transcriptome	Species agnostic *not compatible with Plants
TCR	Human, Mouse	Maps paired TCR sequences together with the T cells' whole transcriptome profiles.
BCR	Human, Mouse	Maps paired BCR sequences together with the B cells’ whole transcriptome profiles.
CRISPR Detect		Enables the screening of extensive guide RNA libraries in up to 1 million cells in a single experiment. This allows the analysis of hundreds to tens of thousands perturbations. The kit offers guide RNA enrichment suitable for 16 distinct sublibraries and is compatible with all three Evercode WT formats.
Gene Select	Human, Mouse	Gene Select is a hybrid capture based enrichment solution allowing research to focus on sequencing only genes of interest.

ESTIMATED SAMPLETHROUGHPUT FOR MGI DNBSEQ G400 AND T7 SEQUENCERS

Based on recommendation of 25K reads per cell Deeper sequencing may be recommended based on library type or study aim

	Number of Samples per Flow Cell
Target Number of Cells	DNBSEQ G400 FCL	DNBSEQ T7 FCL
5,000	14	46
10,000	7	23
20,000	3	11

Single cell experiments 'at scale' - the million cell experiment

"The range of single cell technologies on offer and the improvements in throughput are allowing researchers to rethink their experimental design and run a lot more samples."

Dr. Renee SmithSenior Genomics Scientist

Single Cell RNA (scRNA-seq)

Submit Fresh Cells in sterile microcentrifuge tubes. Cell Submission ~ 1000 – 2000 cells/µl in PBS + 0.04% BSA. Please contact us for more information.

Client prepared libraries should be pooled and submitted in a single tube. A minimum of 10 nM in 100 µl of pooled library should be provided.

Single Cell Sequencing

Offering both microfluidics-based and combinatorial-barcoding single cell methods

DECIDING WHICH SINGLE CELL TECHNOLOGY IS RIGHT FOR YOUR EXPERIMENT.