ThruPLEXTag-seqlibrarieswerepreparedfrom30ngofthe1%HorizoncfDNAstandard.FollowingAgilentSureSelecttargetenrichmentandsequencingtoachieve~10,000Xcoverage,datawasanalyzedwiththeCurioGenomicsbioinformaticsplatform.Usingtherawreads,theEGFRmutation(yellowdot)isobscuredbybackgrounderrors(left).ThemutationisreADIlyidentifiedwhenUMTsareutilizedindataprocessingtocorrectforerrors(right).
Datafromeitherthe30ngHorizoncfDNAlibrary(previousslider)(A)oraThruPLEXTag-seqlibrarypreparedfrom10ngHorizoncfDNAstandard,enrichedwithacustom240KBpanel(NimbleGenSeqCapEZ)andsequencedonaHiSeq2500toachieve~5,000Xcoverage.AnalysiswasperformedusingtheCurioGenomicsbioinformaticsplatform.
Startingwith1to50ngofDNA,ThruPLEXTag-seqKitcreatesindexedlibrariesin3simplesteps:endrepair,adapterligation,andhigh-fidelitylibraryamplification.Nopurificationorsampletransferstepsarerequired.Thestreamlinedworkflowisperformedin2hoursinasingletubeorwell,preventingsamplelossandenhancingpositivesampleidentification.
ThruPLEXtechnologyisa3-stepreactionthatstartswithfragmenteddouble-strandedDNAorcell-freeDNAwhichisrepairedinahighlyefficientprocess.Backgroundisreducedusingdouble-strandedadaptorswithnosingle-strandedtails.Bluntendligationoccurswithhigh-efficiency.Blocked5"endsreduceadapter-adapterligation.
ReachresultsquicklywiththeCurioGenomicscloud-basedsoftware(www.curiogenomics.com)orConnor,anopen-sourcetoolavailablefordataprocessing(https://github.com/umich-brcf-bioinf/Connor).
Guaranteedfor9monthsat-20°Cinaconstanttemperaturefreezer.
Storeat–20°C.
CAT.NO.R400584ThruPLEXTag-seq6S Kit(12reactions,6singleindexes)
CAT.NO.R400585ThruPLEXTag-seq48SKit(48reactions,48singleindexes)
CAT.NO.R400586ThruPLEXTag-seq96DKit(96reactions,96dualindexes)
ThruPLEXTag-seqcontainsallnecessaryreagentsforpreparingindexedIlluminaNGSlibraries,includingoptimizedIllumina-compatIBLeadaptersandindexingreagents.
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