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KAPA Hyper Prep and HiFi Uracil+ Kits Allow for Reduced Representation Bisulfite Sequencing of Ultra-low Amounts of DNA in B Cells

Kapa Biosystems, Inc.

KAPA Hyper Prep and HiFi Uracil+ Kits Allow for Reduced Representation Bisulfite Sequencing of Ultra-low Amounts of DNA in B Cells

17 Dec 2015

This application note provides a robust methodology for RRBS analysis using the streamlined chemistry of the KAPA Hyper Prep Kit and HiFi Uracil+ HotStart DNA Polymerase, and demonstrates its utility in accurately quantifying the DNA methylation status from sample-limiting conditions. Methyl-Seq is a powerful tool for understanding genome-wide methylation with base-pair resolution. Several strategies have been developed to understand the methylation status of a genome, including reduced representation bisulfite sequencing (RRBS). Bisulfite treatment results in a significant decrease in DNA input and quality; therefore, the construction of high-quality libraries and efficient downstream amplification is critical.

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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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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 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.Cancer DiagnosticsThere are a wide variety of diagnostic tests for cancer available, and this range continues to expand as our knowledge of cancer improves. Current diagnostic methods include biopsy, imaging and blood tests for known biomarkers. New methods in research development include liquid biopsies and cancer breathalyzers.AutomationAutomation in laboratories and manufacturing processes enhances efficiency, precision, and scalability by reducing the need for manual intervention. It plays a critical role in improving productivity, minimizing human error, and accelerating workflows in fields like diagnostics, drug development, and industrial testing. Automation technologies include robotic systems, automated liquid handlers, and process control systems that streamline complex tasks and ensure consistent, reproducible results. Explore our peer-reviewed product directory to discover the best automation solutions, compare options, read user reviews, and get prices directly from manufacturers.DNA MethylationMethylationCell BiologyCell biology studies the structure, function, and behavior of cells. Understanding cellular processes is essential for research in areas such as cancer biology, stem cell research, and neurobiology. Techniques like flow cytometry, microscopy, and cell culture enable researchers to explore cellular mechanisms in detail. Browse our peer-reviewed product directory to find the best cell biology tools and equipment, compare products, check customer reviews, and get pricing directly from manufacturers.Developmental BiologyProcess 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.

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KAPA Hyper Prep and HiFi Uracil+ Kits Allow for Reduced Representation Bisulfite Sequencing of Ultra-low Amounts of DNA in B Cells