Transcription
KAPA HiFiAmplify the possibilities
All polymerases are not created equalReliable DNA amplification is critical to the success of many molecular biology methods, from simple cloning and screeningto gene editing and next-generation sequencing (NGS).Typically, DNA polymerases offer EITHER low error rates OR greater speed, range, and inhibitor tolerance. Avoid makingtrade-offs when performing high-fidelity amplification of long or difficult targets. Insist on the combined fidelity andprocessivity ofKAPA HiFi DNA Polymerase: Amplify Your Possibilities.Using Directed Evolution technology (below), we selected for polymerase qualities that increase success rates for a broadrange of applications.Overview of directed evolutionStarting material:gene coding forwild-type polymeraseThe result is an improved,evolved polymerase.The process is repeated,applying additional selectivepressures with each iteration.Upon amplification, only thefittest polymerases self-replicateand are enriched; less fitvariants fail to replicateLibrary of randomvariants createdby mutagenesisVariants transformedinto E. coli, wherethey are expressedVariant-expressing cellscompartmentalized in droplets,then lysed in the presenceof PCR componentsand selective pressureThis resulted in a single, versatile enzyme that combines robust proofreading capabilities with low error rates, high speed,greater sensitivity, and more robust amplification through challenging templates.Whatever your application may be, KAPA HiFi can amplify your successHIGH PROCESSIVITY3 Amplify regions up to 11 kb3 Reduce amplification bias and sequencing bias in AT- or GC-rich regionsLOW ERROR RATES3 Ensure accuracy in expression vectors and gene-editing templates3 Reduce falsely identified SNPs and mutationsHIGH SENSITIVITY3 Detect rare but important molecules in heterogeneous samples3 Capture sequencing data from rare transcripts and low-copy variantsHIGH SPEED3 Save time on workflows that use PCR
Simplify cloning projects with amplification of templates up to 11 kbMany high-fidelity DNA polymerases lack theprocessivity to reliably amplify long templates.KAPA HiFi is not one of them.4.5 kbng505.00.55.0 kb505.07.5 kb0.550 5.09.0 kb0.5505.011.0 kb0.5505.00.5KAPA HiFi readily amplifies templates as long as11 kb from a broad range of template input amounts.Accurately amplify longer templates to reducethe number of PCR reactions in DNA cloningworkflows, ultimately reducing the number of stepsand conserving resources.Amplification of targets from 4.5 kb to 11 kb, starting with 50 ng,5 ng, and 0.5 ng of hgDNA. Each reaction was performed with 35 cycles ofdenaturation, annealing, and extension.Minimize introduction of PCR artifacts when amplifying DNAfor sequencing, cloning, and other applications2.50 x 10-52.00 x 10-5Error RateThe accuracy of KAPA HiFi—fewer than 1 error per3.6 x 106 nucleotides—ensures that PCR productsaccurately represent the original targets, increasingconfidence in downstream results.1.50 x 10-51.00 x 10-55.00 x 10-60.00Taq-basedblendsTaqQ5PhusionKAPAHiFiKAPA HiFi error rates are lower than for other DNA polymerases.The proofreading activity of KAPA HiFi DNA Polymerase outperformsother polymerases, including Taq, Q5 (New England Biolabs), and Phusion(Thermo Scientific).Ensure uniform representation throughout the genomewith robust performance across challenging regionsNo amplif ication2520201510KAPA HiFi3025DepthDepth30151055000.50.60.70.80.5Frag GC0.60.70.8Frag GCPhusion30TruSeq3025252020DepthDepthKAPA HiFi is the enzyme of choice for NGS libraryamplification. Its ability to amplify GC- and AT-richregions with nearly the same efficiency as neutralcontent leads to uniform sequence coverage that isvirtually identical to that obtained with a PCR-freeworkflow. This ensures reliable results andreduces sequencing costs.1515101055000.50.60.7Frag GC0.80.50.60.70.8Frag GCSequence coverage in libraries amplified using KAPA HiFi DNAPolymerase is nearly indistinguishable from that of unamplifiedlibraries. In contrast, GC-rich sequences are under-represented in librariesamplified using Phusion DNA Polymerase (Thermo Scientific) or the TruSeq PCRMaster Mix (Illumina).
Overcome the challenges of amplifying bisulfite-treated DNA—with KAPA HiFi Uracil Bisulfite treatment of methylated DNA creates specifichurdles for DNA polymerases:8Average size (bp) Long stretches of DNA are highly AT-enrichedSize bias360350Concentration (ng/µL) The DNA templates contain uracils, which inhibit theactivity of traditional proofreading polymerasesYield10643403303203103002280KAPA HiFi Uracil is engineered to amplify bisulfitetreated DNA templates with greater efficiency and fidelitythan other polymerases.Whether the converted DNA is used in NGS methods ortraditional cloning, KAPA HiFi Uracil will providehigh yields, low bias, improved quality ofsequencing data, and lower sequencing costs.01214Cycles28016KAPA HiFi Uracil 1214Cycles16Pfu Turbo CxKAPA HiFi Uracil produces higher library yields and minimal sizebias compared to Agilent Pfu Turbo Cx. Human whole genome bisulfitetreated libraries were amplified with 12, 14, or 16 cycles using standard protocolsand the amplified libraries were analyzed using a Bioanalyzer 2100 High SensitivityDNA assay.KAPA HiFi DNA Polymerase drives the success of KAPA library preparation kitsThe high fidelity and processivity of KAPA HiFi DNApolymerase make it the polymerase of choice forproducing NGS libraries that accurately reflect thecomplexity of the original samples, whether the input is:RNA For total RNA-seq, with mRNA enrichment,rRNA depletion, and options for customdepletionDNA For genomic samples, cDNAs, amplicons,degraded samples, targeted sequencing,mechanically or enzymatically sheared inputs,and bisulfite-treated DNAAmplify the success of your projectConsult with our Support & Applications Scientists to: Discuss project-specificworkflow considerations Troubleshoot challenges Develop and install automationscripts and workflows Consult and collaborate to integratenew applications into your lab Receive wet-lab trainingContact us: support.seqls@roche.com
From PCR to NGS, HiFi has you coveredDNA amplification and qPCR R eal-time monitoring oflibrary amplification PCR Cloning Site-directed mutagenesisStandard NGS library preparation for RNA and DNA RNA-seq libraries(total RNA, mRNA,ribodepleted RNA) Whole-genome sequencing (WGS) libraries,when PCR is required N GS target enrichment:amplification of enrichedlibraries for DNAor RNA-seq Amplicon sequencing(e.g., 16S and 18S sequencingfor microbial metagenomics Whole-exome sequencing (WES) librariesEpigenomics and chromatin structure Bisulfite sequencing MIRA TAB-seq Target enrichment for Epi workflows ATAC-seq Hi-C MeDIP-seq WGBS-seqAdditional specialized applications SMART-seq2 CRISPR/Cas9 validation screening CRISPR/Cas9 gene editing CIRCLE-seqThe list is always growing!If your application isn’t listed here, please reach out to us:go.roche.com/hifi
Ordering information250 UKK2501KAPA HiFi HotStart PCR Kit100 UKK2502KAPA HiFi HotStart PCR Kit250 UKK2601KAPA HiFi HotStart ReadyMix1.25 mLKK2602KAPA HiFi HotStart ReadyMix6.25 mLKK2800KAPA HiFi HotStart Uracil Kit10 rxnKK2801KAPA HiFi HotStart Uracil Kit50 rxnKK2802KAPA HiFi HotStart Uracil Kit250 rxnKK2610KAPA HiFi HotStart Library Amp Kit10 rxnKK2611KAPA HiFi HotStart Library Amp Kit50 rxnKK2612KAPA HiFi HotStart Library Amp Kit250 rxnKK2620KAPA HiFi HotStart Library Amp Kit with Primer Mix50 rxnKK2621KAPA HiFi HotStart Library Amp Kit with Primer Mix250 rxnKK2623KAPA Library Amp Primer Mix250 rxnKK2702KAPA HiFi HotStart Real-Time Library Amp Kit250 rxnKK2709KAPA HiFi HotStart Real-Time Library Amp Standards1.5 mLPublished by:Roche Sequencing and Life Science9115 Hague RoadIndianapolis, IN ctollQ100 UKAPA HiFi PCR KitTargetEnKAPA HiFi PCR KitKK2102PKK2101SampleQPack eady LibraryFor more information, contact yourRoche Sequencing representative or visit:go.roche.com/HiFisequencing.roche.comData on file.For Research Use Only. Not for use in diagnostic procedures.KAPA is a trademark of Roche. All other product names and trademarks are the property of their respective owners. 2019 Roche Sequencing and Life Science. All rights reserved.MC-US-03358BRO301001A4392/19
Phusion TruSeq KAPA HiFi. KAPA HiFi DNA Polymerase drives the success of KAPA library preparation kits The high fidelity and processivity of KAPA HiFi DNA polymerase make it the polymerase of choice for producing NGS libraries that accurately reflect the complexity of the original samples, whether the input is: RNA For total RNA-seq, with mRNA enrichment, rRNA depletion, and options for custom .
