Kapa biosystems/KAPA hgDNA Quantification and QC Kit/KK4962/300 x 20 µL reactions,Kapa biosystems,,Kapa biosystems,Kapa,kapabiosystems,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：Kapa biosystems/KAPA hgDNA Quantification and QC Kit/KK4962/300 x 20 µL reactions

产品编号：KK4962

市场价：¥12080.00

场地：美国(厂家直采)

产品分类：分子类>二代测序>靶向测序试剂盒>

联系QQ：1570468124

电话号码：4000-520-616

邮箱： info@ebiomall.com

美元价：$604.00

品牌： Kapa biosystems

公司：Kapa Biosystems.

公司分类：

商品介绍

Description:

qPCRMasterMix(Bio-Rad®)

KAPAhgDNAQuantificationandQCKit

Qualitymatters.

KAPAhgDNAQuantificationandQCKitscontainallthereagentsneededfortheqPCR-basedquantificationandqualityassessmentofhumangenomicDNAsamplespriortoNGSlibraryconstruction.

KitscontainKAPASYBR^®FASTqPCRMasterMix,optimizedforhigh-performanceSYBRGreenI-basedqPCR,aswellasapre-dilutedsetofDNAstandardsandprimerpremixestargetingdifferentportionsofahighlyconservedsingle-copyhumanlocus.Absolutequantificationisachievedwiththeprimerpairdefiningtheshortestfragment,whereastheadditionalprimersareusedtoderiveinformationabouttheamountofamplifiabletemplateintheDNAsample.Qualityscores(orQ-ratios)generatedwiththekitmaybeusedtopredicttheoutcomeoflibraryconstruction,ortailorworkflowsforsamplesofvariablequality,particularlyFFPEDNA.

ThemethodiseasytoautomateandcanbeappliedtoanyprocessorworkflowthatrequiresaccuratequantificationofdiluteDNAsamples,orsamplesthatmaycontainahighproportionofDNAthatisrecalcitranttoPCRamplification.

Downloadour KAPAhgDNAQuantificationandQCDataAnalysisTemplate.

ForResearchUseOnly.Notforuseindiagnosticprocedures.

ProductHighlights

Principle of the hgDNA Quantification and QC assay. A single set of DNA standards is used to generate up to three standard curves using three different primer pairs that amplify targets of 41 bp, 129 bp or 305 bp within a conserved, single-copy human locus. The 41 bp assay is used for absolute quantification of DNA samples. For an assessment of DNA quality, standard curves are generated and samples assayed with the 129 bp and/or 305 bp primer premix(es). Since poor DNA quality has a greater impact on the amplification of longer targets, the relative quality of a DNA sample can be inferred by normalizing the concentration obtained using the 129 bp or 305 bp assay against the concentration obtained from the 41 bp assay. This normalization generates a Q-ratio (with a value between 0 and 1) that is indicative of DNA quality, or the amount of amplifiable material in a DNA sample. Data on file.

Obtainconcentrationandqualityinformationwithasingleassay

AllowsforaccuratequantificationofdiluteDNAsamples
QuantificationwithanadditionalprimerpairprovidesaQ-ratiothatisindicativeofsamplequality

Conversion rates (% input DNA converted to adapter-ligated library) for libraries prepared for targeted re-sequencing on the Illumina platform. Libraries were prepared in duplicate from 80 – 100 ng FFPE DNA of variable quality (orange) or a commercial hgDNA preparation (grey), using the KAPA LTP Library Preparation Kit. FFPE DNA samples were assessed with the KAPA hgDNA Quantification and QC assay prior to Covaris shearing. Q-ratios were as indicated. The amount of fragmented DNA (average size ~180 bp) used for library construction was determined using the Qubit HS dsDNA assay (Life Technologies), whereas adapter-ligated libraries were quantified using the qPCR-based KAPA Library Quantification Kit. Results indicated that conversion rates for FFPE samples were significantly lower than for high-quality DNA. This limits the diversity of FFPE libraries, and necessitates more cycles of library amplification to generate sufficient material for target capture. This results in higher duplication rates and lower target coverage for FFPE libraries. Data on file.

Employqualityscores intheanalysisofNGSlibraryconstructionworkflows

Q-ratioscanprovidevaluableinsightsintothebottlenecksinNGSlibraryconstructionworkflows
ForFFPEsamples,libraryandsequencequalityisprimarilylimitedbyinefficientconversionofinputDNAtoadapter-ligatedlibrary

0.4yieldedlibrariesthatmetminimumsequencequalityrequirements.Dataonfile.">

Q(129/41) ratios for correlate with key sequencing metrics, such as duplication rate. Libraries were prepared manually from 250 ng Covaris-sheared FFPE DNA (n=14) or control DNA, extracted from cultured cells, fresh-frozen tissue or blood (n=6). Samples are arranged on the x-axis (left to right) in order of increasing Q(129/41)-ratio (designated by orange bars for FFPE samples and blue bars for control DNAs). Libraries were constructed with NEBNext reagents (New England Biolabs), whereas KAPA HiFi was used for pre- and post-capture amplification (10 cycles). Target capture was performed with a custom Roche NimbleGen SeqCap EZ panel (300 genes, 0.9 MB). Sequencing (2 x 100 bp) was performed on an Illumina HiSeq. The average duplication rate (grey dots) were 2X higher for FFPE (75% vs. 38% for control samples). FFPE samples with a Q(129/41) ratio >0.4yieldedlibrariesthatmetminimumsequencequalityrequirements.Dataonfile.

0.4yieldedlibrariesthatmetminimumsequencequalityrequirements.Dataonfile.">

Q(129/41) ratios for correlate with key sequencing metrics, such as mean target coverage. Libraries were prepared manually from 250 ng Covaris-sheared FFPE DNA (n=14) or control DNA, extracted from cultured cells, fresh-frozen tissue or blood (n=6). Samples are arranged on the x-axis (left to right) in order of increasing Q(129/41)-ratio (designated by orange bars for FFPE samples and blue bars for control DNAs). Libraries were constructed with NEBNext reagents (New England Biolabs), whereas KAPA HiFi was used for pre- and post-capture amplification (10 cycles). Target capture was performed with a custom Roche NimbleGen SeqCap EZ panel (300 genes, 0.9 MB). Sequencing (2 x 100 bp) was performed on an Illumina HiSeq. The average mean target coverage (grey dots) was 4X higher for control DNA (1,247X, vs. 301X for FFPE samples). FFPE samples with a Q(129/41) ratio >0.4yieldedlibrariesthatmetminimumsequencequalityrequirements.Dataonfile.

PrevNext

Obtainactionabledataforsamplepreparation

Q-ratioscorrelatewithkeysequencingmetricssuchasduplicationratesandmeantargetcoverage
FFPEsampleswithaQscore>0.4yieldlibrariesofacceptablequalitywhenprocessedinstandardsamplepreparationsfortargetcapture

Applications:

Applications

FFPEsamples
Free-circulatingDNAfromplasmaorserum
Samplesobtainedbylaser-capturemicrodissectionoffresh,frozenorFFPEtissue
Forensicsamples
Cellscollectedbyflowcytometry
Anyotherlimitingorpreciousclinicalsample

KitSpecificationsandContents/Storage:

KitSpecificationsandContents/Storage

Kitscanbestoredforupto12months at-20˚C.

Complete(MasterMix)kitsincludeKAPASYBRFASTqPCRMasterMix(2X),PrimerPremix(41bp,129bpand305bp,10X)andasetof5DNAStandards.PrimerPremixesandDNAStandardsarealsosoldseparately.

Components

Specifications

CompatIBLePlatform

AllNGSplatforms

CompatibleSamples

HumangenomicDNA

Samplesources

FFPEtissueCellscollectedbylaser-capturemicrodissectionFlow-sortedcellsFree-circulatingDNAfromplasmaorserumForensicsamplesClinicalsamples

Standardcurveconcentrationrange

2.5ng/µL–10pg/µLor3,080–12copies

SequencingApplications

WholeGenomeSequencingWholeExomeSequencingTargetedSequencing(custompanels)AmpliconSequencing

品牌介绍

Kapabiosystems

Kapa是世界上著名的生物技术公司，总部位于美国。特色产品：Kapa2G快速PCR、实时荧光定量PCR、Kapa长片段DNA多聚酶、Kapa高保真DNA多聚酶。

We are constantly evolving better science by applying the latest advances in genomics and synthetic biology to help reduce costs, simplify workflows and improve results. Our core protein engineering technologies enable us to tailor enzymes to suit specific applications in next-generation DNA and RNA sequencing, DNA amplification and molecular diagnostics. Biomedical research is no longer limited by the constraints of native enzyme structure or function.

产品列表：

No.	品牌	货号	名称	规格
1	kapa	KK4824	Complete kit (Universal)	500 x 20 µL reactions
2	kapa	KK4854	Complete kit (optimized for Roche® LightCycler 480)	500 x 20 µL reactions
3	kapa	KK4873	Complete Kit (ROX Low)	500 x 20 µL reactions
4	kapa	KK4973	qPCR MasterMix (ROX Low ) and Primer Premix only	500 x 20 µL reactions
5	kapa	KK4827	Complete kit (Universal)	500 x 20 µL reactions
6	kapa	KK4847	Complete kit (Bio-Rad®)	500 x 20 µL reactions
7	kapa	KK4924	qPCR MasterMix (Universal) and Primer Premix only	500 x 20 µL reactions
8	kapa	KK4944	qPCR MasterMix (Bio-Rad) and Primer Premix only	500 x 20 µL reactions
9	kapa	KK4813	Library Quantification Primer Premix	500 µL
10	kapa	KK2502	HotStart PCR Kit, with dNTPs	250U
11	kapa	KK2102	With dNTPs	250U
12	kapa	KK2602	HotStart ReadyMix, 500 x 25 µL reactions	6.25 mL
13	kapa	KK4701	100 x 20 µL reactions (Universal)	1 ml
14	kapa	KK4705	100 x 20 µL reactions, ROX (ABI Prism®)	1 ml
15	kapa	KK4709	100 x 20 µL reactions, with fluorescein (Bio-Rad® iCycler™)	1 ml