Targeted Exome and Gene Panel Analysis from Low Input and FFPE DNA using Hybridization Capture for Cancer Genome Studies

2 Aug 2016

Hybridization-based target enrichment techniques coupled with Next-Generation Sequencing (NGS) provide a useful and cost-efficient means to study disease specific target regions including whole exomes and gene panels. Clinical samples may be limited in input and of compromised quality due to formalin fixation, however most current NGS library preparation methods require 50-100 ng high quality DNA for such studies. This application note presents an efficient method, which enables high quality target enrichment and variant calling from inputs as low as 1-25 ng.

SureSelect Focused Exome

Agilent Technologies

The SureSelect Focused Exome is a highly targeted design that enables analysis of only disease-associated targets, enabling superior coverage of disease-associated regions even on benchtop sequencers. This design provides 20 or more reads for 95% of targets at 1.5Gb (100x) of sequencing and 98% of targets with 3Gb (200x) of sequencing.

(0)

SureSelect Target Enrichment System Kit

Agilent Technologies

Agilent's SureSelect market leading platform provides a complete portfolio of catalog and custom products, providing the flexibility you need, for your discovery to follow-up studies. Agilent™s SureSelect Target Enrichment System is a highly efficient hybrid selection technique for optimizing next-generation sequencing. It is available for the Illumina GA and HiSeq, Applied Biosystems SOLiD, and 454 Life Sciences FLX and GS Junior NGS systems, including paired or single-end protocols.SureSelect Target Enrichment System Kit Features: Custom design your kit through eArray. Highly automatable, easy design process. Kit sizes can be chosen to accommodate very small projects, up to thousands of samples. Contents include all reagents and customer-specified libraries.

(1)

SureSelect DNA Panels

Agilent Technologies

Agilent SureSelect DNA panels are predefined designs which focus on targeted gene sets for specific applications. Agilent offers two human panels, the SureSelect Human Kinome panel and SureSelect X-Chromosome panel. The SureSelect Human DNA Kinome panel targets a comprehensive set of kinases and kinase related genes for enrichment, targeting over 500 kinases and 612 genes. The SureSelect X-Chromosome panel targets all of the genes on the X chromosome. • SureSelect Kinome kits are available for targeting either DNA (genomic regions) or RNA (transcripts) • Efficient workflows for post-capture (XT) and pre-capture (XT2) pooling – (XT2 - pool up to 16 samples per enrichment)

(0)

Links

Tags

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.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.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.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.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. :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.Target-EnrichmentGene ExpressionLibrary PreparationLibrary preparation is a critical step in sequencing workflows, where DNA or RNA samples are converted into libraries for high-throughput, next generation sequencing. This step ensures accurate results and minimizes biases. Explore library preparation kits in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.DNA AmplificationDNA Amplification is a technique used to amplify a single or multiple copies of DNA or mRNA by orders of magnitude. The most common method employed is PCR, but other options exist that eliminate the need for thermocycling.Cancer ResearchCancer research aims to understand the mechanisms of cancer development and progression to improve prevention, diagnosis, and treatment. From molecular biology to clinical trials, research spans a wide range of disciplines, including immunotherapy, targeted therapies, and drug discovery. Explore the best cancer research products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Genomic HybridizationFFPEFormalin-fixed paraffin-embedded (FFPE) tissues are commonly used in medical research, particularly in histology and cancer studies. The preservation process allows long-term storage of tissue samples for later analysis. Explore FFPE research products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.