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ThruPLEXPlasma-seqKitgeneratedqualitylibrarieswithhighdiversityandalownumberofduplicatesandunmappedreads.Cell-freeDNAwasextractedfrom3plasmasamples,andlibrarieswerepreparedattheamountsindicatedasmeasuredbyqubit®.TheamountofmononucleosomalDNAineachsampleasmeasuredbytheBioanalyzer®correspondedto0.09ng,0.62ng,and15.44ng.PooledlibrariesweresequencedonanIlluminaNextSeq®500asapaired-endrunwith17Mto25Mreadsperlibrary,Duplicationrateswerecalculatedafterdown-samplingthedatato17Mreadsperlibrary.
ThruPLEXPlasma-seqKitlibrarieswerecapturedathighefficiencyandgenerateddatawithdeepcoverageofthekinomeformutationdetection.Librarieswerepreparedfrom3plasmasamplesatinputamountsof5ng,6.5ng,and10ngintriplicate,andtargetedsequencingwascarriedoutonanIlluminaMiSequsingsamplesenrichedwiththeClearSeqHumanDNAKinomePanelforSureSelectXT2.Onaverage,5Mreadsweregeneratedperlibrary.Selectedbasesweresuccessfullycapturedbasesthatwereinorwithin250bpofthebaits.
ThruPLEXPlasma-seqKitprovidedthemostreproducibleandunbiasedGCcoverageacrossthehumangenome.ThruPLEXlibrariesshowedminimalvariABIlityacross9individualplasmasamplestested.Librarieswerepreparedfromcell-freeDNAisolatedfrom1mLofplasmasamplesandsequencedonanIlluminaNextSeq500.FourseparateplasmasampleswereusedtoconstructtheNEBNextUltralibraries.
IlluminaNGSlibrarieswerepreparedwith5ngofcell-freeDNAisolatedfrompooledplasmasamplesusingThruPLEXPlasma-seqKit.Low-passWGSwasconductedonaHiSeqwithapproximately12.5Mreadsperlibrary.(DatacourtesyoftheKarolinskaInstitute,Sweden).
Startingwith1to30ngofcell-freeDNA,ThruPLEXPlasma-seqKitcreatesindexedlibrariesin3simplesteps:endrepair,adapterligation,andhigh-fidelitylibraryamplification.Nopurificationorsampletransferstepsarerequired.Thestreamlinedworkflowisperformedin2hoursinasingletubeorwell,preventingsamplelossandenhancingpositivesampleidentification.
ThruPLEXPlasma-seqKittechnologyisa3-stepreactionthatisoptimizedforcell-freeDNA.Cell-freeDNA(1ngto30ng)isfirstrepairedinahighlyefficientprocess.Backgroundisreducedusingdouble-strandedadapterswithnosingle-strandedtails.Blunt-endligationoccurswithhigh-efficiency.Blocked5’endsreduceadapter-adapterligation.Backgroundisfurtherreducedbydestroyingunusedadaptersafterligation.Ahigh-fidelityamplificationcompletesthereactiontogenerateindexedIlluminalibraries.
Guaranteedfor9monthsat-20°Cinaconstanttemperaturefreezer.
Storeat–20°C.
CAT.NO.R400490ThruPLEXPlasma-seq12S Kit(12reactions,12singleindexes)
CAT.NO.R400491ThruPLEXPlasma-seq48SKit(48reactions,48singleindexes)
CAT.NO.R400492ThruPLEXPlasma-seq96DKit(96reactions,96dualindexes)
ThruPLEXPlasma-seqcontainsallnecessaryreagentsforpreparingindexedIlluminaNGSlibraries,includingoptimizedIllumina-compatibleadaptersandindexingreagents.
SouthernBiotech/Goat Anti-Human Lambda-Alexa Fluor® 555/2070-32/1.0 mg
vectorlabs/Biotinylated Aleuria Aurantia Lectin (AAL)/B-1395/1 mg
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Mouse IgG)/PK-6102/1 kit
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Universal)/PK-6200/1 kit
vectorlabs/Unconjugated Musa Paradisiaca (Banana) Lectin (BanLec)/L-1410/5 mg
GiottoBiotech/Calmodulin N60D/5 mg/G02CLM60cn