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蚂蚁淘在线 / 品牌中心 / Kapa biosystems / Kapa生物系统/Kapa文库扩增试剂盒/KK2621/250 x 50µL反应

Kapa生物系统/Kapa文库扩增试剂盒/KK2621/250 x 50µL反应

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.
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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是世界上著名的生物技术公司。总部位于美国波士顿, 公司的主要理念是运用分子和遗传学技术来打造新一代的生物科学和医学诊断产品。Kapa公司目前主要是在“直接进化（directed evolution technology platform）”的技术平台上来筛选和生产高品质的生物酶。目前生物医药领域所使用的大部分生物酶都只是天然存在形式，即使有些酶经过基因工程改造，但是由于酶分子自身的结构和功能限制，要想得到真正满足科研工作人员需要的产品，却是一件费时费力的事情，而且常常达不到预期的要求。“直接进化”技术正是针对这一现实情况，运用遗传，基因工程，生物信息学等技术来在分子水平上设计真正满足不同实际要求的生物酶。通过创新，Kapa为广大的科研人员提供了一系列拥有多项技术的产品，不仅能够满足普通科研需要，同时为高通量，基因芯片等高新技术的普及创造了条件。 运用“直接进化”技术，Kapa目前已经开发