Single-cell RNA Sequencing

CD Genomics provides robust transcriptome research service down to single-cell input levels in high-quality samples These global gene expression patterns in single cells already have dramatically advanced cell biology.

The Introduction of Single-cell RNA Sequencing

Cells are the basic unit of life and each cell is unique. The ability to reveal complex cellular events in biological systems is critical to a better understanding of cellular contributions during development or in disease progression. Gene expression research at the single-cell resolution on samples composed of mixed cell populations allows for deep insight into c into the transcriptome complexity of diverse cell types.

CD genomics offers best-in-class tools for library preparation and sequencing from a single cell, a few cells, and ultra-low inputs of RNA. Combining the robust cDNA synthesis technology with Illumina next-generation sequencing and analysis technologies, we offer reliable data of the highest quality to study cell-to-cell transcriptome heterogeneity

Single-cell RNA Sequencing Workflow

The advent of cell sorting/partitioning technologies, such as flow cytometry and microfluidics, has made it possible to capture single cells, and the DNA or RNA of single cells is amplified for single-cell sequencing. The general workflow for single-cell RNA sequencing is outlined below.

Techniques Description

Fluidigm C1 Single-Cell mRNA Workflow. With Fluidigm C1 system, We provide single-cell transcriptome profiling service at an optional scale. C1 can rapidly and reliably capture and process individual cells. The steps in Integrated C1 Single-Cell mRNA Seq workflow include fluidics circuits (IFCs) to capture cells, convert polyA+ RNA into full-length cDNA, and perform universal amplification of the cDNA. With the customizable microfluidic circuits, C1 enables seamless transition from identifying critical cell populations to generate sequencing libraries for transcript 3′ End Counting, full-length mRNA sequencing, DNA sequencing, epigenetic analysis, micro-RNA expression profiling and more.

SMART-seq Ultra Low Input RNA for sequencing. The SMART technology provides full-length transcript information, enabling analysis of transcript isoforms, gene fusions, point mutations, etc. The kit employing this technology enables users to perform cDNA synthesis directly from 1–1,000 intact cells or ultra-low input total RNA with high reproducibility and accurate representation of GC-rich transcripts. Full-length cDNA libraries produced with this kit are compatible with Illumina platforms.

Service Specifications

	Sample requirements 96-well plates of live cells suspension are recommended for this procedure. Frozen cells in specifically frozen lysates can also be used as starting material. >100 pg extracted RNA, OD260/280 ratio is between 1.8 to 2.0
	Sequencing Flexible service options include HiSeq X and MGI DNBSEQ-T7/DNBSEQ-G400
	Bioinformatics Analysis Raw data quality control Statistics of sequencing depth and coverage SNP/InDel/SV/CNV calling Annotation and statistics Pathway enrichment analysis Population genetics analysis More data mining upon your request

Deliverables

• The original sequencing data
• Experimental results
• Data analysis report
• Details in Single-Cell Sequencing for your writing (customization)

CD Genomics's Single-Cell Sequencing conference focuses on the links between cell variation in tissues and organ function and further elucidates the origins of diseases. If you have additional requirements or questions, please feel free to contact us.

References

1. Wu, AR et al. Quantitative assessment of single-cell RNA-sequencing methods. Nature Methods. 2014 January; 11(1): 41–46.
2. Picelli, S et al. Full-length RNA-seq from single cells using Smart-seq2.Nature Protocols. 2014 Jan;9(1):171-81.