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蚂蚁淘在线 / 品牌中心 / Kapa biosystems / Kapa biosystems/KAPA Library Amplification Kits/KK2621/250 x 50 µL reactions

Kapa biosystems/KAPA Library Amplification Kits/KK2621/250 x 50 µL reactions

价格
￥13000.00
货号：KK2621
浏览量：127
品牌：Kapa biosystems
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商品描述

Description:

Withprimermix

KAPALibraryAmplificationKits

ThegoldstandardforNGSlibraryamplification.

KAPALibraryAmplificationKitscontainKAPAHiFiDNAPolymerase,anovelenzymeengineeredforultra-highfidelityandrobustnessusingKapa’sdirectedevolutiontechnologyplatform.

KAPAHiFihasbecometheenzymeofchoiceforNGSlibraryamplificationduetoitsABIlitytoamplifycomplexDNApopulationswithhighfidelity,highefficiency,decreasedPCRduplicationratesandverylowbias.*ThisresultsinlowerduplicationratesandimprovedcoverageofGC-andATrichregions,promoters,lowcomplexityandotherchallengingregionsinallNGSlibraryconstructionworkflowsrequiringlibraryamplification.

Inadditiontothestandardlibraryamplificationformulation,KAPAHiFiisavailableinauniquereal-timeformulation,forapplicationsthatdemandprecisecontroloverlibraryamplification.Auracil-tolerantvariant,KAPAHiFiUracil+isalsoavailableforthehigh-efficiency,high-fidelity,low-biasamplificationoflibrariesconstructedfrombisulfite-treatedDNA.

KAPALibraryAmplificationKitsforIllumina® platformsnowalsoincludeanoptimallyformulatedLibraryAmplificationPrimerMix.

*Dataonfile.

ForResearchUseOnly.Notforuseindiagnosticprocedures.

ProductHighlights

Coverage depth vs GC content for indexed Illumina TruSeq libraries prepared from 1 µg M. tuberculosis (65% GC) gDNA. Libraries amplified with KAPA HiFi HotStart or the Illumina TruSeq PCR Master Mix are compared to an equivalent unamplified library. Data was generated by paired-end sequencing (2 x 75 bp). After filtering and aligning read pairs to reference sequences, 250 000 read pairs were randomly sampled for each genome, and scatter plots of mean sequence coverage depth vs. GC content were generated by analyzing 250 bp windows. GC-rich sequences were under-represented in libraries amplified with the TruSeq PCR Master Mix. In contrast, library amplification with KAPA HiFi resulted in coverage distribution across the range of GC-content that is almost indistinguishable from that of the unamplified control. Data on file.
Key sequencing metrics for libraries prepared from 1 µg human gDNA with the KAPA HTP Library Preparation Kit (green) or TruSeq DNA Sample Prep Kit (orange), for multiplexed exome capture using the SeqCap EZ Human Exome Library v3.0 (Roche Nimblegen). KAPA HiFi only required fewer cycles of pre- and post-capture amplification (7 and 14, respectively), as compared to the 8 cycles and 18 cycles performed libraries generated and amplified with Illumina enzymes. The PCR duplication rate for libraries amplified KAPA libraries were significantly lower, leading to improved coverage metrics. Data on file.
PrevNext

Achievethelowestamplificationbiasandduplicationrates

  • Loweramplificationbiasresultinimprovedrepresentationofalllibraryfragmentsandsequenceregions
  • High-efficiencyamplificationleadstofeweramplificationcyclesandlowerPCRduplicates
  • Lessadditionalnext-generationorSangersequencingneededtocompletegenomes
The first exons of many genes have a high GC content, and are therefore difficult to amplify. Regions of these exons are therefore typically underrepresented in whole exome sequence data. Pre-capture amplification with the evolved, low-bias KAPA HiFi DNA Polymerase results in significantly improved coverage of GC- and AT-rich regions, and other sequence elements that are difficult to amplify. Data courtesy of The Broad Institute. Data on file.

ImprovecoverageofGC-andAT-richregions

  • LoweramplificationbiasimprovescoverageuniformityofGC-andAT-richregions,promoters,lowcomplexityandotherchallengingregions
  • ImprovedoverallcoverageandcoverageuniformityobservedonbothIllumina andIonTorrent™ sequencingplatforms
Real-time, high fidelity amplification of multiple libraries with the KAPA Real-Time Library Amplification Kit. Twenty libraries, spanning a ~64-fold concentration range (equivalent to 6 cycles), were simultaneously amplified and terminated after 14 cycles. Fourteen of the twenty libraries fall within the targeted amplification range (the fluorescence range defined by the Real-Time Amplification Standards 1¬–3). The remaining six libraries could either be used as is, noting that they may be outside the optimal concentration range, or they could be re-amplified individually or in high- or low-concentration groups. Data on file.

Exerciseprecisecontroloverlibraryamplification

  • KAPAReal-TimeAmplificationKitsallowforlibraryamplificationtobeobservedinrealtime
  • Terminateamplificationattheoptimalpointforindividualsamples
  • Optimizelibraryamplificationparametersforhigherthroughputworkflows
Error rates of Taq DNA polymerase and polymerases typically used in NGS library amplification. The error rates of proofreading DNA polymerases such as Q5 (New England Biolabs), Phusion (Thermo Scientific) and the evolved KAPA HiFi DNA Polymerase is 50 – 100 times lower than that of Taq. The error rate of KAPA HiFi was confirmed by deep sequencing on the 454 platform, and is 1 error per 3.54 x 106 nucleotides incorporated. Data on file.

AmplifyNGSlibrarieswithindustry-leADIngfidelity

  • KAPAHiFiwasengineeredusingdirectedevolutiontohaveenhancedproofreading(3′-5′exonuclease)activity
  • Industry-leadingfidelityconfirmedby454sequencing

Applications:

Applications
  • AmplificationofIllumina libraries(DNAandRNAsequencing)
  • Real-timeamplificationoflibrariespreparedfromlowinput,ChIPDNAandotherprecioussamples
  • Pre-andpost-captureamplificationintargetedcaptureworkflows

KitSpecificationsandContents/Storage:

KitSpecificationsandContents/Storage

Kitscanbestoredforupto12months at-20˚C.

KitsincludeKAPAHiFiHotStartReadyMix(2X),aconvenientPCRmastermixcontainingKAPAHiFiHotStartDNAPolymerase,KAPAdNTPs,reactionbuffer,andMgCl2atafinalconcentrationof2.5mM.Kitswithprimersarealsoavailable.

Kitsforreal-timelibraryamplificationincludeareal-timeversionoftheKAPAHiFiHotStartReadyMix,aswellasfluorescentstandardstomonitorlibraryamplification.

Components

LAK_Component Chart

Specifications

Spec
Description
CompatIBLePlatform
IlluminaHiSeq,MiSeq,NextSeq,andGAIIx
LibraryType
DNA,RNA,andbisulfite-treatedDNA
StartingMaterial
AnyNGSlibrarythatrequiresamplification
SequencingApplications
WholeGenomeSequencingWholeExomeSequencingTargetedSequencing(custompanels)RNA-SeqChIP-SeqAmpliconSequencing
Kapa biosystems表观遗传分析基于阵列的方法和基于下一代测序（NGS）的方法都用于研究表观遗传修饰。有三种基于NGS的常见方法可用于表观遗传分析：甲基序列，ChIP序列和ATAC序列。Methyl-seq   用单核苷酸分辨率研究基因组的甲基化状态。该方法采用亚硫酸氢盐处理，可将胞嘧啶残基转化为尿嘧啶，而甲基化残基则保持不变。已经开发了几种甲基序列策略，包括全基因组亚硫酸氢盐测序（WGBS）和简化表示的亚硫酸氢盐测序（RRBS），这丰富了CpG岛。ChIP-seq将染色质免疫沉淀（ChIP）与NGS结合使用，以鉴定整个基因组中与DNA相关的蛋白质的结合位点，通常用于绘制组蛋白修饰和转录因子。该方法依靠靶向抗体的选择来富集与特定蛋白质结合的目标DNA片段。ATAC-seq是用于转座酶可及的染色质测序的一种测定方法，可确定染色质可及性的区域并绘制DNA结合蛋白的图谱，以鉴定活性启动子，增强子和其他顺式调控元件。该方法通过生成具有50,000个细胞的测序文库，已经改变了基因调控的分析方法。由于表观遗传学分析通常涉及超低输入DNA，因此从有限的材料构建高质量文库至关重要。罗氏测序解决方案提供了许多用于表观遗传工作流程的目标富集，文库制备和优化文库质量的解决方案。SeqCap®Epi靶标富集试剂盒可通过单碱基分辨率富集用于DNA甲基化评估的靶标。的  KAPA HyperPrep套件  理想地适合于这两个芯片起和甲基SEQ应用，因为它使接头连接的文库和扩增低偏压的更高的产率。这意味着更高的文库多样性，更低的重复率和更统一的覆盖率，尤其是对于低输入样本而言。对于甲基序列研究，  KAPA HiFi Uracil + 由于对尿嘧啶残基具有耐受性，HotStart DNA聚合酶对于亚硫酸氢盐转化的文库的扩增至关重要。KAPA HiFi HotStart ReadyMix可用于扩增ATAC-seq和ChIP-seq库，从而改善序列覆盖范围并减少偏差。