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Improve your NGS and Enable Novel Applications with Engineered DNA Polymerases from Kapa Biosystems

Kapa Biosystems, Inc.
27 Mar 2016

Using a number of case studies, this poster illustrates that engineered DNA polymerases are capable of providing better sequence coverage and are enabling novel and challenging NGS applications. The vast majority of PCR applications utilize a small number of wild-type DNA polymerases; improvements to existing applications and the development of novel PCR applications have primarily been realized via innovative changes to all of the reaction components except the polymerases themselves. Kapa Biosystems have engineered a new generation of improved DNA polymerases such as the KAPA HiFi. These are capable of robust amplification of diverse targets spanning a wide range of sizes and GC-content, and KAPA HiFi has demonstrated production of high yields with remarkably little amplification bias when used for NGS library amplification.

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KAPA HiFi Uracil+

Kapa Biosystems, Inc.

KAPA HiFi DNA Polymerase provides high fidelity next-generation sequencing library amplification with the highest yield, lowest bias, and most uniform sequence coverage available. These characteristics are retained in KAPA HiFi Uracil+ DNA Polymerase, a modified version of KAPA HiFi engineered to tolerate uracil residues. The new enzyme is particularly well suited for applications employing bisulfite DNA conversion, which typically produces low concentrations of AT-rich DNA. KAPA HiFi HotStart Uracil+ ReadyMix (2X) is a ready-to-use cocktail containing all components required for PCR, except primers and template.   Features: Higher yields and reduced size bias More uniform sequence coverage Improved representation of AT-rich sequences   Applications: Methylation analysis – amplification of bisulfite-converted DNA for sequencing Amplification of damaged DNA samples Prevention of false-positive results due to carryover amplicon contamination

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KAPA HiFi PCR Kits

Kapa Biosystems, Inc.

KAPA HiFi DNA Polymerase is a novel, single-enzyme system that exhibits industry-leading performance when compared with other high fidelity polymerases and polymerase blends. KAPA HiFi has been engineered to have an increased affinity for DNA without the need for accessory protein domains. The intrinsic high processivity of KAPA HiFi results in significant improvements to yield, sensitivity, speed, target length, and the ability to amplify difficult templates (e.g AT- and GC-rich). The error rate of KAPA HiFi PCR Kits is 100X lower than wild-type Taq DNA polymerase.   Features: Achieve the highest fidelity Increased processivity, strong proofreading activity, and optimized buffer system result in superior accuracy The error rate of KAPA HiFi is 100X lower than Taq, 40X lower than polymerase blends, 3X lower than Pfu Ultra and 2X lower than Phusion Improve performance on GC- and AT-rich templates Achieve higher success rates across targets up to 84% GC content Higher yields when amplifying AT-rich templates Amplify longer targets with greater sensitivity  High-fidelity PCR of long and complex genomic templates Improved sensitivity High speed allows significantly shorter reaction times for long range PCR   Applications: High-fidelity PCR Amplification of DNA fragments for cloning and protein expression or genomic characterization Site-directed mutagenesis Next-generation sequencing library amplification

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KAPA DNA Library Preparation Kits for Illumina

Kapa Biosystems, Inc.

KAPA DNA Library Preparation Kits for Illumina sequencing platforms contain evolved and optimally formulated enzymes that enable the highest overall coverage from the least amount of total sequencing. Kits are optimized to achieve significantly higher library yields and contain KAPA HiFi for high-fidelity, high-efficiency and low-bias library amplification. This ensures the highest library and sequence data quality, particularly from low input and difficult samples such as FFPE and ChIP DNA. Features: Achieve greater molecular complexity Higher yields of adapter-ligated library translates to higher molecular complexity Fewer cycles of amplification are needed, which results in lower PCR duplication rates PCR-free workflows possible from inputs ≥100 ng Reduce amplification bias and improve coverage KAPA HiFi reduces PCR bias Improves coverage uniformity of GC- and AT-rich regions, promoters, low complexity and other challenging regions Create high-quality libraries from challenging samples High-quality whole exome sequencing from libraries prepared using as little as 10 ng human genomic DNA High success rates with 250 ng FFPE or less Routine library construction from ≥100 pg ChIP DNA Automate with pre-validated scripts PerkinElmer Sciclone NGS and NGSx Beckman Coulter Biomek FXp and FX dual-pod Agilent Technologies NGS Workstation Option B Eppendorf epMotion 5075 TMX   Applications: Whole Genome Sequencing ChIP Sequencing Whole Exome Sequencing Amplicon Sequencing Targeted Sequencing (Capture) Methyl Sequencing

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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.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.qPCRQuantitative PCR (qPCR) is a molecular biology technique used to amplify and measure the amount of a specific DNA or RNA target. It plays a crucial role in gene expression analysis, pathogen detection, and quantifying DNA or RNA in various applications. Real-time PCR provides precise, quantitative results and is widely used in research, diagnostics, and clinical studies. Browse our peer-reviewed product directory to find the best qPCR solutions; compare products, check reviews, and get pricing directly from manufacturers.Library GenerationLibrary generation refers to the construction of NGS libraries from RNA and DNA sources.Library 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.Process OptimizationProcess optimization involves improving the efficiency, cost-effectiveness, and quality of processes in various industries, including manufacturing, pharmaceuticals, and biotechnology. Explore process optimization solutions in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Genome SequencingGenome sequencing involves determining the complete DNA sequence of an organism's genome. It provides crucial information about genetic variations, mutations, and diseases. Advances in sequencing technologies, such as next-generation sequencing (NGS), have accelerated research in genomics, diagnostics, and personalized medicine. Explore genome sequencing tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.RT qPCRReal-time quantitative PCR (RT-qPCR) is a technique used to measure gene expression and quantify DNA or RNA in real-time. It is essential in genomics, diagnostics, and biotechnology research. Explore RT-qPCR systems in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Real-Time PCRReal-time PCR (RT-PCR) is a technique used to amplify and simultaneously quantify nucleic acids in real time. It is a key tool in gene expression analysis, pathogen detection, and diagnostics. RT-PCR allows for the detection of specific RNA or DNA sequences with high sensitivity and specificity. Browse our peer-reviewed product directory to find the best Real-Time PCR systems; compare products, check reviews, and get pricing directly from manufacturers.

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Improve your NGS and Enable Novel Applications with Engineered DNA Polymerases from Kapa Biosystems