ProductHighlights:
TheTruSeqCustomAmpliconLowInputLibraryPrepKitisascalableampliconsequencingsolutionthatdeliverssensitiveandspecificresultsfrombothlow-inputandformalin-fixed,paraffin-embedded(FFPE)DNAsamples.Itoffers:
- Accuratevariantdetectionfromaslowas10ngofinputgenomicDNA(gDNA)andchallengingFFPEsamples
- Fullysupported,optimizedworkflowsolutionincludingsimpledataanalysis
- Automation-friendlyworkflowthatcanbecompletedin6hours,withonly3hoursofhands-ontime
LowDNAInputandFFPECompatibility
TheTruSeqCustomAmpliconLowInputLibraryPrepKitisafullycustomizable,amplicon-basedassayfortargetedresequencingstartingfromaslowas10ngofgenomicDNA(gDNA).Thisscalableassayallowsresearcherstocapturemultipletargetsofinterestsimultaneouslyandsequenceupto1536ampliconsinasinglepool,usingasinglereaction.ThislibraryprepkitofferstheflexibilitytoaccommodateFFPEsamples,suchaspreservedtumortissue.
ConfidentAssayDesign
DesigntargetedTruSeqCustomAmpliconoligonucleotideprobeswithDesignStudio,afree,easy-to-useonlinetoolthatprovidesoptimizedcoverage.
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AssessFFPESampleQualitybeforeSequencing
TheTruSeqFFPEDNALibraryPrepQCKitusesasimpleqPCRreactiontodetermineDNAqualityandprovideguidanceonsequencingparameters.Thisstepensuresthatonlysamplesthatwillachievethenecessarysequencingmetricsareprepared,conservingresourcesthatmightbeusedonpotentiallylow-quality,unrecoverablesamples.TheresultsoftheQCstepdeterminetherecommendedamountofinputDNA.TheTruSeqFFPEDNALibraryPrepQCKitcanbebundledwiththeTruSeqCustomAmpliconLowInputLibraryPrepKittomaximizelabbudgets.
ForFFPEapplications,wesuggestdesigningyourprojectwith150bpor175bpampliconssinceFFPEDNAishighlydegraded.TruSeqCustomAmpliconLowInputcannotbeusedwithapplicationsrequiring425bpamplicons.
SpeciesCompatibility
TheTruSeqCustomAmpliconLowInputLibraryPrepKitsupportshuman,mouse,rat,andbovinesamples.DesignStudiodesignalgorithmsarealsoavailableformaize,rice,pig,dog,soybean,chicken,andsheep;however,assayperformancecannotbeguaranteedastheresultingassayshavenotbeentested.Inaddition,IlluminaConciergeservicescandesigncustomassaysand/orprovideassayoptimizationsupportforanyspeciesofinterest.
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Findanup-to-datelistofhigh-throughputautomationvendorswithroboticsystemscompatIBLewiththislibrarypreparationkit
Specifications:
| AssayTime | 1day |
| Hands-OnTime | 4hours |
| InputQuantity | 50ngRNA,50nghigh-qualitytotalRNA,≥200ngFFPEtotalRNA;Recommendedquantitymayvarywithexpressionlevel,targetplexity,andsamplequality |
| ContentSpecifications | Choosefrom400,000+pre-designedtargetedRNA-Seqassays.Oraddcontenttoafixedpanelorpreviouslydesignedcustompanel. |
| Multiplexing | Upto384samplespersequencingrun |
| MechanismofAction | Amplification |
TruSeqCustomAmpliconLibraryPrepProtocol:
TheTruSeqCustomAmpliconassayisasimpleandstreamlinedmethodforcapturingandamplifyingtargetedregionsofinterest.SeamlessAmpliconSequencingWorkflow:
TheTruSeqCustomAmpliconLowInputLibraryPrepKitispartofanintegrated,fullysupportedworkflowforampliconsequencingthatguidesresearchersfromdesignthroughdataanalysis.Illuminatechnicalandfieldspecialistshelpensurerapidresolutionandminimizepotentiallaboratorydowntime.RobustVariantDetectionatLowDNAInput:
TheTruSeqCustomAmpliconLowInputLibraryPrepKitdemonstrateshighconcordancebetweenexpectedandobservedvariantfrequenciesfor10nghigh-qualityFFPEDNA.ReferenceDNAsampleswerepreparedfollowingtheTruSeqCustomAmpliconLowInputworkflowwiththeTruSeqAmplicon-CancerPanelprimerpool,andsequencedontheMiSeqSysteminreplicatesof4.VariantswerecalledusingMiSeqReporterSoftware.R2valuesareshown.
