A Standardized, Automated Approach for Exosome Isolation and Characterization Using Beckman Coulter Instrumentation

7 Jun 2016

This application note describes a workflow using automated Biomek methods for centrifugation layering and fractionation, total RNA extraction, and cDNA amplification and clean-up for next generation sequencing. NGS results are reported on benign and cancerous colon cell lines.

Biomek® 4000 Laboratory Automation Workstation

Beckman Coulter Life Sciences

Biomek Liquid Handlers enable you can spend less time on manual process and more time on powerful discoveries. Beckman Coulter Life Sciences is a leader when it comes to providing solutions and support that cover the full spectrum of the NGS sample preparation workflow with reagents and automation. Throughput options allow you to choose the right solution for your laboratory.

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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.Cell Lines Stem Cells and Primary CellsPrimary cell cultures, established cell lines and stem cells are vital for <i>in vitro</i> and <i>ex vivo</i> experimentation. High-quality cells, optimized for your applications, alongside optimized cell substrates, growth medium and supplements, are critical for experimental success. Explore a range of cells suitable for your applications, including isogenic cell lines, competent cells, induced pluripotent stem cell (iPSC)-derived cell lines, fungal/bacterial/mammalian cell lines, stem cells and cancer cell lines. Find the best cells for your research in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.CentrifugesCentrifuges are used to separate particulates suspended in a liquid via the application of a centrifugal force including cells, macromolecules, nanoparticles and precipitates. Centrifuges come in three sizes: microcentrifuges, benchtop and floor-standing centrifuges, depending on the volume and number of samples that require separation. When choosing a centrifuge, consider its maximum separation power indicated by its relative centrifugal force (RCF) value. Low force centrifuges reach a maximum RCF of 8,000 g, high-force centrifuges can reach maximum RCF values of 80,000 g. For extremely high forces, consider ultracentrifuges (up to 800,000 g). Refrigerated and vacuum centrifuges also known as concentrators are available for more specialized applications. A range of volumes are available for centrifuge tubes and microfuge tubes, depending on sample sizes. Find the best centrifuges in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Sample ManagementSample management systems include sample storage devices such as freezers and plate storers, sample environment enclosures and sample organization, retrieval and sorter systems. Useful system features include high-throughput, automation, robotic arms, automated liquid handling and associated database systems. Accessories in sample management include barcode scanners, heat sealers and tubes.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving 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.NanotechnologyNanotechnology, or nanotech, is an engineering technique using molecular scale functional systems. Applications of nanotechnology include medicine and medical devices, electronics, air and water purification, food science and energy production.Gene Expression and Molecular CloningMolecular cloning is a set of techniques that utilizes vectors to transfer recombinant DNA into host cells and is an essential tool for investigating the expression of genes and proteins in bacterial or mammalian cells. A variety of vectors optimized for gene cloning and expression in a range of host organisms are available, alongside competent cells for genetic replication. Here, you can explore a range of molecular tools, high-quality genomic and cDNA libraries, premade clones, transformation and transfection reagents and mutagenesis or gene expression detection assays and expression arrays. Find the best gene expression and molecular cloning products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.RNA InterferenceRNA interference (RNAi) uses siRNA or miRNA for transcriptional silencing, gene knockdown and regulation of gene expression. RNAi requires chemical synthesis, introduction of DNA vectors into cells, an assay of RNAi effects and RNAi quantification or analysis. Consider target sequence selection, reagent preparation, controls, high specificity and effectiveness and low non-specific gene knockdown.BiomarkersBiomarkers are biological markers which can be measured and evaluated to indicate a biological state. The use of biomarkers in research and diagnosis can indicate a normal or disease state or drug response of cells / tissues. Biomarkers include genetic markers, cell surface markers such as antigens, antibodies or receptors and secreted molecules such as cytokines. An assay system is required for identification of biomarkers. :Robotic InstrumentsRobotic instruments can be used for high-throughput automation of many lab processes. Such processes use instruments for assays like cell based assays and ELISA, for sample preparation like shakers, centrifuges and incubators, and for analysis such as sequencing and western blot analyzers. Useful features of robotic instruments include speed, reproducibility, barcode readers, software and automation.Software PlatformsSoftware platforms are useful for various stages of laboratory experiments from data collection to data storage and processing. For instance lab software is available for system control, data management, data analysis and qualification / validation.Clinical GeneticsMolecular Genetics covers the analysis of hereditary genetic disease and chromosomal abnormalities. Genetics can be analysed using DNA, RNA, and protein microarrays, PCR, RT PCR and DNA sequencing. Genetic equipment includes genetic workstations, thermal cyclers, cooling blocks and electrophoresis products. Diagnostic kits are used for DNA / RNA extraction and purification.