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DNA扩增、二代测序、分子诊断
℡ 4000-520-616
℡ 4000-520-616
Kapa biosystems/KAPA hgDNA Quantification and QC Kit/KK4963/300 x 20 µL reactions
产品编号:KK4963
市  场 价:¥12080.00
场      地:美国(厂家直采)
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:
Kapa biosystems/KAPA hgDNA Quantification and QC Kit/KK4963/300 x 20 µL reactions
商品介绍

Description:

qPCRMasterMix(optimizedforRoche®LightCycler480)

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

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

Spec
Description
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


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