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蚂蚁淘在线 / 品牌中心 / Kapa biosystems / Kapa biosystems/KAPA Library Quantification Kits/KK4924/500 x 20 µL reactions

Kapa biosystems/KAPA Library Quantification Kits/KK4924/500 x 20 µL reactions

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￥9460.00
货号：KK4924
浏览量：127
品牌：Kapa biosystems
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商品描述

Description:

qPCRMasterMix(Universal)andPrimerPremixonly

KAPALibraryQuantificationKits

forNext-GenerationSequencing

Ourstandardsdon’tchange.Neithershouldyours.

KAPALibraryQuantificationKitscontainallthereagentsneededfortheaccurate,reliableandreproducIBLeqPCR-basedquantificationofNGSlibrariespriortopoolingforcaptureorflowcellamplification.KitscontainKAPASYBR®FASTqPCRMasterMix,optimizedforhigh-performanceSYBRGreenI-basedqPCR.TheengineeredKAPASYBRFASTDNAPolymerasecontainedintheMasterMixamplifiesGC-andAT-richDNAfragmentsofdifferentlengthswithsimilarefficiency,makingittheonlyqPCRMasterMixcapableofaccurateqPCR-basedquantificationofNGSlibraries.*Thepre-dilutedsetofDNAStandardscontainedineachkitiscarefullyquality-controlledtoensurelot-to-lotconsistencyandavoiddatadriftovertime.OptimizedPrimerPremixesensureoptimalPCRefficiency.

KitswithDNAStandardsandPrimerPremixesforIllumina® andIonTorrent™sequencers areavailable.Pleasereferencethe InstrumentCompatibilityChart forguidanceoncompatibleplatforms.

NEW!ImproveexperimentaldesignswithourTechnicalGuideforIlluminaPlatforms


*Dataonfile.

ForResearchUseOnly.Notforuseindiagnosticprocedures.

ProductHighlights

qPCR-based KAPA Library Quantification Kits reduce variability in cluster density compared to Agilent Bioanalyzer. Human exome samples were prepared using Nimblegen target capture. All preparations were performed in a 96-well plate format on a liquid handling system. Average number of clusters per tile are shown for paired-end, 76-bp Illumina GAIIx runs. Data courtesy of the University of Washington. Data on file.
Compared to spectrophotometric, fluorimetric, and electrophoretic methods, qPCR only quantifies adaptor-ligated and amplifiable library molecules. The ligation of adaptor sequences (pink and blue) to library DNA molecules (grey) results in a mixed population of molecules without the correct adaptor configuration. Electrophoresis and spectrophotometry measure total nucleic acid concentrations, whereas optimal cluster density or template-to-bead ratio depends on the appropriate concentration of PCR-amplifiable DNA molecules. Only qPCR-based library quantification methods count library molecules with the correct adaptor sequence configuration. Data on file.
PrevNext

AccuratelyquantifyPCR-competentsequencingtemplates

  • qPCRonly“counts”thoselibrarymoleculesthatcanbesequenced
  • Moreconsistentclusterdensitiesortemplate-to-beadratiosenableoptimalutilizationofsequencingresources
Equimolar pooling of indexed libraries. Eleven indexed Illumina TruSeq libraries were quantified by qPCR using the KAPA Library Quantification Kit, and then combined to achieve equal final concentrations in two separate pools for multiplexed sequencing on different flow-cell lanes. The eleven libraries ranged ~11-fold in concentration from 0.67 pM to 7.65 pM, while representation of each index varied between 90% and 127% of expected assigned reads per lane. Data on file.

Improvepoolingformultiplexedsequencing

  • LibraryconcentrationsdeterminedbyqPCRaremorereliablecomparedtoothermethods*
  • TheengineeredKAPASYBRFASTqPCRMasterMixenablesaccuratepoolingoflibrarieswithextremeGC-contentsorlongerinserts*
Nine human DNA libraries and two microbial DNA libraries (Rhodococcus sp. ~70% GC and Staphylococcus sp. ~35% GC) were used to compare quantification results obtained with distinct lots (”Lot 1” and “Lot 2”), and distinct sets of reagents from the same lot (”Set 1” and “Set 2”) of KAPA Library Quantification Kits for Illumina platforms. Data on file.

Eliminatedriftwithcarefullyquality-controlledDNAStandards

  • PredilutedDNAStandardseliminatequantificationerrorsassociatedwithpreparationofstandardcurvesusingin-housestandards*
  • KAPADNAStandardsundergostrictqualitycontroltoensurelot-to-lotconsistencyandeliminatedatadriftovertime
KAPA Library Quantification methods (96- and 384-well) format are available for the Sciclone NGS and NGSx workstation (PerkinElmer) and other instruments used in high-throughput NGS sample preparation pipelines.

Improvethroughputwithautomation

  • Libraryquantificationassayiscompatiblewith96-and384-wellformat
  • Librarydilution,reactionsetupanddataanalysiscanbeautomatedforHTPpipelines
  • LearnMore aboutKAPAAutomatedSolutions

*Dataonfile.

RelatedProducts

Normalizinglibrariesforpoolingandclusteramplification?CheckouttheseKapaNGSproductstoimproveyourworkflowandresults:

Sample QC

KAPAhgDNAQuantificationandQCKit

Library Amplification

KAPALibraryAmplificationKits

Library Preparation

KAPAHyperPrepKits

KAPA Stranded mRNA-Seq Kits

KAPAStrandedmRNA-SeqKits

Applications:

Applications
  • Quantificationoflibrariespriortopoolingformultiplexedsequencing
  • QuantificationofindividuallibrariesorlibrarypoolspriortoclusteramplificationoremPCR
  • Quantificationoflibrariespriortoandafterhybridizationcapture
  • Qualitycontrol,optimizationandtroubleshootingoflibraryconstructionprocessesorworkflows

KitSpecificationsandContents/Storage:

KitSpecificationsandContents/Storage

Kitscanbestoredforupto12months at-20˚C.

CompletekitsincludeKAPASYBRFASTqPCRMasterMix(2X),PrimerPremix(10X)andasetof6DNAStandards.ReagentsforqPCR(KAPASYBRFASTqPCRMasterMixandPrimerPremix)andDNAStandards(1–6)arealsosoldseparately.Wherenoted,MasterMixescontaininstrument-specificreferencedyes,whiletheUniversalkitincludesROXHighandROXLow(both50X)separately.

Components–Illumina®Platform

Components–IonTorrent™Platform

LQK-Ion_Component Chart

Specifications

Spec
Description
CompatiblePlatforms
IlluminaHiSeq,NextSeq,MiSeq,andGAIIxIonTorrentProtonandPGM454GSFLX+andGSJuniorSOLiD5500series
LibraryType
Any
StartingMaterial
AnyNGSlibraryreadyforclusteramplificationoremPCR
Standardcurveconcentrationrange
20–0.0002pMforIlluminalibraries83–0.00083pMforIonTorrentand454libraries10–0.0001pg/µLforSOLiDlibraries
SequencingApplications
WholeGenomeSequencingWholeExomeSequencingTargetedSequencing(custompanels)RNA-SeqChIP-SeqAmpliconSequencingMethyl-Seq
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。