After cDNA derived from antibody-derived tags (ADTs) is separated from the mRNA-derived cDNA by size selection,the cell-surface-protein library and gene-expression library are prepared independently, and could be sequenced together with different sample indexes. The DNA-barcoded antibodies are incubated with single-cell suspensions under conditions comparable to flow cytometry staining protocols, then the unbound antibody is removed by washing. This kind of antibody is suitable for both platforms. The antibody-oligo complexes are incubated with single-cell suspensions using conditions comparable to staining protocols used in flow cytometry, after which cells are washed to remove unbound antibodies and processed for scRNA-seq. conjugated well-characterized monoclonal antibodies to oligos that were designed to (1) contain a poly-A tail, making it compatible with poly-dT capture RNA-seq-based systems, (2) contain a specific barcode sequence for unique antibody identification. Prior to single-cell genomics, defining cell-surface proteins, which are often reliable indicators of cellular activity and function, is critical to understanding the unique characteristics of the various cell types. Simultaneously Measure Proteins and RNA at a Single-cell Level We compared them in terms of mechanisms, operations and some scRNA-seq results obtained with them.Ģ.1.2. In this review, we compare two of the most successful commercial single-cell sequencing platforms: BD Rhapsody and 10x Genomics Chromium (Fig. More recently, microfluidics platforms and droplet-based methods have enabled massively parallel sequencing of mRNA in large numbers of individual cells. These SCS techniques include the detection of DNA mutations, copy-number variants (CNVs), DNA-protein binding, RNA splicing, and the measurement of mRNA expression. Sequencing the DNA of individual cells can give information about mutations carried by a small population of cells, for example in cancer, while single-cell RNA-seq can investigate transcriptomic cell-to-cell variation, revealing new cell types, and providing insights into developmental processes and transcriptional stochasticity. The rapidly evolving single-cell sequencing (SCS) technology has become a powerful method to resolve complex tissue compositions and delineate convoluted cellular development. When analyzing sequencing data from bulk tissue samples, typically which are composed of millions of cells, it is difficult to resolve cell to cell variations and identify cells that may play an important role in disease progression. Generate consistent and reproducible single cell gene expression results across experiments, across users, and even across multiple sites.The Cell is the fundamental unit of an organism. Use this automated solution to get sequencing-ready libraries every day and reclaim valuable lab time.Ĭonsistent and reproducible library prep: Chromium Connect integrates components customized for the 10x Genomics single cell workflow, automation-specific reagents and consumables, and an easy-to-use touchscreen computer. Save time and resources: Chromium Connect reduces errors from manual pipetting and is estimated to shorten your hands-on time from more than 8 hours to less than an hour. Run your assays with the confidence that technical variation is minimized with this validated instrument. Reduce technical variability: Chromium Connect delivers consistent and reproducible single cell data across experiments and across users. With Automation for Single Cell Immune Profiling workflows, you can automate your 5’ gene expression and V(D)J amplification workflows to examine cellular heterogeneity of the immune system and T- and B-cell receptor diversity, at scale. One system, many single cell assays: With Automation for Single Cell Gene Expression, you can automate your 3’ gene expression workflow to explore cellular heterogeneity, discover novel targets and biomarkers, and unravel complex cellular processes, with increased productivity. LC-MS innovation: Improve analytical accuracy Which microbioreactor modules are best for you Which solid support is right for your oligo synthesis?
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