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蚂蚁淘在线 / 品牌中心 / Kapa biosystems / Kapa biosystems/KAPA RNA HyperPrep Kits with RiboErase (HMR)/KK8560/24 libraries

Kapa biosystems/KAPA RNA HyperPrep Kits with RiboErase (HMR)/KK8560/24 libraries

价格
￥43200.00
货号：KK8560
浏览量：127
品牌：Kapa biosystems
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商品描述

Description:

KAPARNAHyperPrepwithRiboErase

KAPARNAHyperPrepKitswithRiboErase(HMR)

Single-DayRNA

TheKAPARNAHyperPrepKitswithRiboErase(HMR)utilizenovelchemistrythatenablesthecombinationofenzymaticstepsandfewerreactionpurifications,resultinginatrulystreamlinedsolutionforthepreparationofhigh-qualityrRNA-depletedRNA-seqlibraries.Byutilizingatargetedenzymaticmethodfordepletion,ourworkflowenablesefficientreductionofribosomalRNA(rRNA)andamorecompleterepresentationofthetranscriptome,includingprecursormRNAsandnon-codingRNA(ncRNA).Thestrand-specificworkflowisflexIBLe–supportinglibraryconstructionfromlower-inputamountsanddegradedsamples.KitscontainallreagentsrequiredforrRNAdepletionandlibrarypreparation,withtheexceptionofKAPAAdapters(availableseparately).Benefitsinclude:

  • single-daylibraryconstruction,inclusiveofrRNAdepletion
  • reducedhands-onandoveralltimethroughfewerenzymaticandreactioncleanups
  • flexibleinputof25ng–1µgtotalRNAfromhuman,mouse,orratspecies*
  • highersuccessrateswithlowerinputanddegradedsamples
  • maintainover99%strandspecificity*
  • KAPAPureBeadsincludedforreactionpurifications

NEW!KAPADual-IndexedAdapterKitsarenowavailable. FormoreinformationonKAPAAdapterKits,scrolldowntotheOrderingsection,ordownloadtheKAPAAdapterandBeadCalculator.DownloadourAdapterandBeadCalculator

 

*Dataonfile.
ForResearchUseOnly.Notforuseindiagnosticprocedures.

ProductHighlights

The KAPA RNA Hyper Prep Kit with RiboErase (HMR) results in a reduction in the total number of reads wasted due to both PCR duplicates and alignment to rRNA loci in comparison to the TruSeq and NEBNext kits. Libraries were generated in quadruplicate with 25 ng of high-quality Universal Human Reference (UHR) RNA using the manufacturers’ standard recommendations per workflow, where possible. Sequencing was performed using an Illumina HiSeq® 2500 in High Output mode with v4 chemistry and 2 x 100 bp read length. Reads aligning to rRNA were removed and paired reads were randomly subsampled to 14M for comparative analyses, including marked duplicates. Data on file.
Better utilize sequencing capacity. (B) Libraries were generated in quadruplicate with 25 ng (rRNA depletion) and 50 ng (mRNA capture) of high-quality UHR RNA using the manufacturers’ standard recommendations per workflow, where possible. Data on file.
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Sequencewhatmatters

  • WastefewerreadsduetothecombinationofrRNAcarryoverandPCRduplicates
  • Identifymoreuniquetranscriptsandgeneswithequivalentsequencing
Improved coverage uniformity. For the top 1000 transcripts, the KAPA RNA HyperPrep workflows result in more even coverage across transcript lengths, as assessed by both a normalized coverage plot and coverage coefficient of variation (CV), in comparison to competitor workflows. (A) Libraries were generated with 25 ng (rRNA depletion) and 50 ng (mRNA capture) of high-quality UHR RNA using the manufacturers’ standard recommendations per workflow, where possible. Data on file.
Improved coverage uniformity. For the top 1000 transcripts, the KAPA RNA HyperPrep workflows result in more even coverage across transcript lengths, as assessed by both a normalized coverage plot and coverage coefficient of variation (CV), in comparison to competitor workflows. (B) Improved coverage of GC-rich regions (outlined in red) of the YBX1 (rRNA depletion) and SLC2A4RG (mRNA capture) genes was demonstrated using the Kapa workflows. Data on file.
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Achievesuperiorcoverageuniformity

  • Obtainmoreuniformdistributionofreadsacrosstranscripts
  • ImprovecoverageofdifficultGC-richtranscripts
Identify more genes and transcripts. With equivalent sequencing, the KAPA RNA HyperPrep Kit with RiboErase (HMR) in comparison to competitor workflows identified more genes and transcripts. Data on file.
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Generatehigh-qualitylibrariesfromdegradedsamples

  • Inputaslittleas25ngwithFFPEsamples,dependingontotalRNAquality
  • Achievelowduplicationratesandhighlyefficient,reproduciblerRNAremovalwithdegradedsamples
  • Identifymoreuniquetranscriptsandgeneswithequivalentsequencing
High-level of agreement between paired FFPE and fresh frozen expression data, in transcripts per million (TPM). Pearson correlation coefficients show a higher degree of agreement with the KAPA RNA Hyper Prep Kit with RiboErase in comparison to the TruSeq Stranded Total RNA Library Prep with Ribo-zero Gold and NEBNext Ultra Directional Library Preparation with the rRNA Depletion Kit. Libraries were generated in duplicate using 100 ng inputs of paired FFPE-derived and fresh frozen breast tumor total RNA samples using the manufacturers’ standard recommendations per workflow. Sequencing was performed using an Illumina HiSeq® 2500 in High Output mode with v4 chemistry and 2 x 100 bp read length. Reads aligning to rRNA were removed and paired reads were randomly subsampled to 14M for comparative analyses, including marked duplicates. Data on file.
High-level of agreement between paired FFPE and fresh frozen expression data, in transcripts per million (TPM). Pearson correlation coefficients show a higher degree of agreement with the KAPA RNA Hyper Prep Kit with RiboErase in comparison to the TruSeq Stranded Total RNA Library Prep with Ribo-zero Gold and NEBNext Ultra Directional Library Preparation with the rRNA Depletion Kit. Libraries were generated in duplicate using 100 ng inputs of paired FFPE-derived and fresh frozen breast tumor total RNA samples using the manufacturers’ standard recommendations per workflow. Sequencing was performed using an Illumina HiSeq® 2500 in High Output mode with v4 chemistry and 2 x 100 bp read length. Reads aligning to rRNA were removed and paired reads were randomly subsampled to 14M for comparative analyses, including marked duplicates. Data on file.
High-level of agreement between paired FFPE and fresh frozen expression data, in transcripts per million (TPM). Pearson correlation coefficients show a higher degree of agreement with the KAPA RNA Hyper Prep Kit with RiboErase in comparison to the TruSeq Stranded Total RNA Library Prep with Ribo-zero Gold and NEBNext Ultra Directional Library Preparation with the rRNA Depletion Kit. Libraries were generated in duplicate using 100 ng inputs of paired FFPE-derived and fresh frozen breast tumor total RNA samples using the manufacturers’ standard recommendations per workflow. Sequencing was performed using an Illumina HiSeq® 2500 in High Output mode with v4 chemistry and 2 x 100 bp read length. Reads aligning to rRNA were removed and paired reads were randomly subsampled to 14M for comparative analyses, including marked duplicates. Data on file.
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Achievereliableresultswithdegradedinputs

  • Attainahigh-degreeofexpressioncorrelationbetweenpairedFFPEandfreshfrozensampleswhichprovidesincreasedconfidenceinsequencedataaccuracy

RelatedProducts

Areyousequencinglow-input,FFPEorhighqualityDNA? RNA? CheckouttheseKapaNGSproductstoimproveyourworkflowandresults:

KAPA mRNA HyperPrep Kits

KAPAmRNAHyperPrepKits

KAPA Stranded mRNA-Seq Kits

KAPAStrandedmRNA-SeqKits

Library Amplification

KAPALibraryAmplificationKits

Library Preparation

KAPAHyperPrepKits

Library Quantification

KAPALibraryQuantificationKits

Applications:

Applications
  • Wholetranscriptome
  • Geneexpressionanalysis
  • Detectionofgenefusions,isoforms,andotherstructuralvariants
  • Noveltranscriptidentification,includingnoncodingtranscripts
  • SNVdiscovery

KitSpecificationsandContents/Storage:

KitSpecificationsandContents/Storage

EnzymesandbuffersforrRNAdepletion,CDNAsynthesis,andlibrarypreparationcanbestoredforupto10monthsat-20°C.KAPAPureBeadscanbestoredforupto10monthsat4°C.(USonly)

KitscontainHybridizationBuffer,HybridizationOligos(HMR),DepletionBuffer,RNaseH,DNaseBuffer,DNase,Fragment,PrimeandEluteBuffer(2X),1stStrandSynthesisBuffer,KAPAScript,2ndMarkingBuffer,2ndStrandSynthesis&A-TailingEnzymeMix,LigationBuffer,DNALigase,PEG/NaClSolution,KAPAPureBeads,LibraryAmplificationPrimerMix(10X),andKAPAHiFiHotStartReadyMix(2X).

RNAHyperRibo_Component Chart

Specifications

Spec
Description
CompatiblePlatform
0616
LibraryType
RNA
StartingMaterial
High-qualityanddegradedtotalRNA
InputAmount
25ng–1µg
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。

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