
Description:
qPCRMasterMix(ROXLow)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

Obtainconcentrationandqualityinformationwithasingleassay
- AllowsforaccuratequantificationofdiluteDNAsamples
- QuantificationwithanadditionalprimerpairprovidesaQ-ratiothatisindicativeofsamplequality

Employqualityscores intheanalysisofNGSlibraryconstructionworkflows
- Q-ratioscanprovidevaluableinsightsintothebottlenecksinNGSlibraryconstructionworkflows
- ForFFPEsamples,libraryandsequencequalityisprimarilylimitedbyinefficientconversionofinputDNAtoadapter-ligatedlibrary
- 0.4yieldedlibrariesthatmetminimumsequencequalityrequirements.Dataonfile.">
- 0.4yieldedlibrariesthatmetminimumsequencequalityrequirements.Dataonfile.">
Obtainactionabledataforsamplepreparation
- Q-ratioscorrelatewithkeysequencingmetricssuchasduplicationratesandmeantargetcoverage
- FFPEsampleswithaQscore>0.4yieldlibrariesofacceptablequalitywhenprocessedinstandardsamplepreparationsfortargetcapture
Applications:
Applications- FFPEsamples
- Free-circulatingDNAfromplasmaorserum
- Samplesobtainedbylaser-capturemicrodissectionoffresh,frozenorFFPEtissue
- Forensicsamples
- Cellscollectedbyflowcytometry
- Anyotherlimitingorpreciousclinicalsample
KitSpecificationsandContents/Storage:
KitSpecificationsandContents/StorageKitscanbestoredforupto12months at-20˚C.
Complete(MasterMix)kitsincludeKAPASYBRFASTqPCRMasterMix(2X),PrimerPremix(41bp,129bpand305bp,10X)andasetof5DNAStandards.PrimerPremixesandDNAStandardsarealsosoldseparately.
Components
Specifications
- SpecDescription
- CompatIBLePlatformAllNGSplatforms
- CompatibleSamplesHumangenomicDNA
- SamplesourcesFFPEtissueCellscollectedbylaser-capturemicrodissectionFlow-sortedcellsFree-circulatingDNAfromplasmaorserumForensicsamplesClinicalsamples
- Standardcurveconcentrationrange2.5ng/µL–10pg/µLor3,080–12copies
- SequencingApplicationsWholeGenomeSequencingWholeExomeSequencingTargetedSequencing(custompanels)AmpliconSequencing
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先谢了
2 单细胞测序是指单个细胞水平上对基因组进行测序。
3、靶向测序步骤为 样品准备、探针/引物设计、目标序列捕获、文库制备、上机测序、数据分析。
4、单细胞的分离--DNA/RNA的提取和扩增(全基因组扩增和全转录组扩增)---测序以及后续的分析和应用。
相同:使用二代测序
不同:样本处理,文库构建,检测的区域,数据分析流程
2.pgem-3zf (+) 双链dna对照模板 0.2g/l,试剂盒配套试剂。
3.m13(-21)引物 tgtaaaacgacggccagt,3.2μmol/l,即3.2pmol/μl,试剂盒配套试剂。
4.dna测序模板 可以是pcr产物、单链dna和质粒dna等。模板浓度应调整在pcr反应时取量1μl为宜。本实验测定的质粒dna,浓度为0.2g/l,即200ng/μl。
5.引物 需根据所要测定的dna片段设计正向或反向引物,配制成3.2μmol/l,即3.2pmol/μl。如重组质粒中含通用引物序列也可用通用引物,如m13(-21)引物,t7引物等。
6.灭菌去离子水或三蒸水。
7.0.2ml或和0.5ml的pcr管 盖体分离,pe公司产品。
8.3mol/l 醋酸钠(ph5.2) 称取40.8g naac·3h2o溶于70ml蒸馏水中,冰醋酸调ph至5.2,定容至100ml,高压灭菌后分装。
9.70%乙醇和无水乙醇。
10.naac/乙醇混合液 取37.5ml无水乙醇和2.5ml 3mol/l naac混匀,室温可保存1年。
11.pop 6测序胶 abi产品。
12.模板抑制试剂(tsr) abi产品。
13.10×电泳缓冲液 abi产品。
14.abi prism 310型全自动dna测序仪。
15.2400型或9600型pcr仪。
16.台式冷冻高速离心机。
17.台式高速离心机或袖珍离心机。向左转|向右转

