The Illumina Bio-Rad Single-Cell Sequencing Solution

1 Oct 2017

Single-cell RNA sequencing provides deep insight into cell function, disease progression and therapeutic efficacy by allowing researchers to differentiate individual cells in heterogeneous populations. This application note presents the Illumina Bio-Rad single-cell sequencing solution as a robust, scalable and user-friendly workflow. It was developed by the industry leaders in sequencing and Droplet Digital™ technologies, enabling transcriptome profiling of hundreds to tens of thousands of single cells in a single experiment.

ddSEQ™ Single-Cell Isolator

Bio-Rad

The ddSEQ Single-Cell Isolator is part of the Illumina® Bio-Rad® Single-Cell Sequencing Solution, a platform that brings Bio-Rad's Droplet Digital™ technology and Illumina's market-leading sequencing expertise to single-cell gene expression studies. The ddSEQ Single-Cell Isolator works with the Illumina® Bio-Rad® SureCell™ WTA 3' Library Prep Kit for the ddSEQ™ System. This technology utilizes disposable microfluidic cartridges to coencapsulate single cells and barcodes into subnanoliter droplets, where cell lysis and barcoding occur. RNA-Seq libraries are subsequently prepared and sequenced. Analysis is conducted via BaseSpace. Features and Benefits Process hundreds to tens of thousands of cells per day Sensitive and unbiased characterization of transcriptional signatures Scalable solution to accommodate a wide range of experimental designs Agnostic to mammalian cell size, enabling unbiased profiling of diverse cell populations End-to-end single-cell sequencing solution with simple, powerful data analysis Applications and Uses Assess cell-to-cell heterogeneity and identify new sub-populations Map cell trajectories Dissect transcriptional mechanics Infer gene regulatory networks Neurology research — normal brain function, neurological disease and psychological disorders Immunology research — B Cell and T Cell subunit functional analyses Cancer research — studies of cellular expansion of clonal cells in tumors and mutations that lead to differences in gene expression and drug response Stem cell research — studies to better understand development and differentiation, including reverse engineering tissues Included Components ddSEQ Test Cartridges ddSEQ Cartridge Holder ddSEQ Single-Cell Isolator Power supply and power cord User manual and product documentation

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Cell / Tissue CultureCell culture or tissue culture is used to study the biology of cells or tissues and to isolate cellular products in an environment which can be manipulated and well defined. Accurately control your culture environment with bioreactors or culture incubators, bind your cells to a surface or together with an extracellular matrix. Distinguish cell types with differential media or proliferate cells with certain characteristics using selective media. Enrich your media with supplements such as growth factors, sera and vitamins. Find the best cell and tissue culture products, kits and equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Genome AnalysisGenomics, the study of genomes, includes functional genomics, evolutionary genomics and comparative genomics. There are many genomic technologies such as DNA sequencing of whole genomes, computational biology and bioinformatics. DNA and nucleic acids must be isolated and concentrated from cells for analysis with kits, automated analyzers and software. Other useful technologies for studying genomics include PCR, microarrays and electrophoresis.OligonucleotidesOligonucleotides are small nucleic acid polymers, usually less than 20 bases in length. Oligonucleotides can be made via enzymatic cleavage or more commonly by chemical synthesis with polymerases. Their use includes FISH, southern blots, microarrays and as primers in PCR. High fidelity synthesis kits and detection systems are available for easy production and detection, respectively.ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.PCR and Thermal CyclingPolymerase chain reaction (PCR) kits and thermal cyclers are used for the in vitro amplification of DNA permitting subsequent analysis and experimental procedures. Explore a range of high-quality polymerase, primers and nucleotides or simplify your workflow with a PCR mastermix. Find reverse transcription PCR (RT-PCR) and cDNA synthesis kits for RNA products and libraries. Quantitatively measure the amplification of DNA with real-time PCR (qPCR) and droplet digital PCR (ddPCR) kits and systems, and discover automated PCR setup solutions to increase throughput. Alternative DNA amplification methods also include recombinase polymerase amplification (RPA) kits. Find the best PCR kits and thermal cyclers and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA / RNA Extraction and PurificationPurified DNA and RNA are required for numerous downstream molecular biology applications. Consequently, the importance of high-quality DNA/RNA extraction and purification equipment cannot be underestimated. Many purification kits are available and are typically optimized for nucleic acid type and source, including plasmid DNA, genomic DNA, mRNA, RNA and viral nucleic acid purification kits. Automated extraction and purification of nucleic acids can be implemented with magnetic bead separator instruments or high-throughput purification workstations. Find the best DNA/RNA extraction and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA SequencingDNA sequencing, such as sanger sequencing, is a biological technique that determines the precise order of nucleotide bases in a fragment or template of DNA. DNA sequencers and genetic analyzers are based on capillary electrophoresis, where labeled DNA fragments are electrophoretically separated by size as they migrate through a polymer. Find the best DNA sequencing products, including DNA sequencing kits, genomic libraries and genetic identity kits in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Next Generation SequencingNext-generation sequencing (NGS), also known as whole-genome sequencing, high-throughput sequencing and massive parallel sequencing, produces and analyses thousands to millions of nucleotide sequences at once. Sequencing systems operate via varying technologies depending on the manufacturer, including sequencing by synthesis, ligation, pyrosequencing, ion semiconductor and single-molecule real-time sequencing. For NGS, library preparation is paramount to successful sequencing. In this section, explore a range of library preparation kits, from targeted, amplicon-based or hybridization-based kits including epigenomic, transcriptomic and genomic workflows to fragmentation kits. Find the best next-generation sequencing products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Droplet Digital PCRDroplet digital PCR (ddPCR) is a method of quantitative PCR that partitions a sample into droplets, allowing precise detection of low-abundance targets. It is commonly used in gene expression analysis and mutation detection. Explore ddPCR systems in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Single Cell AnalysisSingle-cell analysis involves studying individual cells to gain insights into their behavior, gene expression, and function. This approach is valuable in cancer research, stem cell biology, and immunology. Explore single-cell analysis products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.SequencingSequencing refers to determining the order of nucleotides in a DNA or RNA molecule. This process is essential in genomics, diagnostics, and evolutionary biology. Techniques like next-generation sequencing (NGS) and Sanger sequencing provide high-throughput and precise data for analyzing genomes and transcriptomes. Browse our peer-reviewed product directory to find the best sequencing tools and reagents, compare products, check reviews, and get pricing directly from manufacturers.Transcriptomics