《Single-Step Capture and Sequencing of Natural DNA forDetection of BRCA1 Mutations 天然DNA的單步捕獲和測序 BRCA1基因突變的檢測》由會員上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。

1、Downloadedfromgenome.cshlp.orgonAugust13,2018-PublishedbyColdSpringHarborLaboratoryPressSingle-StepCaptureandSequencingofNaturalDNAforDetectionofBRCA1MutationsJohnF.Thompson*,JeffreyG.Reifenberger*,EldarGiladi,KristenKerouac,JaimeGill,ErikHansen,AvakKahvejian,PhilipKapranov,TravisKnope,Dor

2、onLipson,KathleenE.Steinmann,andPatriceM.MilosHelicosBioSciencesCorporationOneKendallSquare,Building200Cambridge,MA02139*Theseauthorscontributedequallytothepublication.Correspondingauthor:JohnF.Thompsonjthompson@helicosbio.comPhone:617-264-1888FAX:617-264-1700Runningtitle:BRCA1mutationsvia

3、single-stepcapture/sequencingKeywords:NextGenerationSequencing,cancer,targetedcapturePage1Downloadedfromgenome.cshlp.orgonAugust13,2018-PublishedbyColdSpringHarborLaboratoryPressAbstractGenetictestingfordiseaseriskisanincreasinglyimportantcomponentofmedicalcare.However,testingcanbeexpensiv

4、ewhichcanleadtopatientsandphysicianshavinglimitedaccesstothegeneticinformationneededformedicaldecisions.TosimplifyDNAsamplepreparationandlowercosts,wehavedevelopedasysteminwhichanygenecanbecapturedandsequenceddirectlyfromhumangenomicDNAwithoutamplification,usingnoproteinsorenzymespriortose

5、quencing.Extractedwhole-genomeDNAisacousticallyshearedandloadedinaflowcellchannelforsingle-moleculesequencing.Geneisolation,amplification,orligationisnotnecessary.AccurateandlowcostdetectionofDNAsequencevariantsisdemonstratedfortheBRCA1gene.Disease-causingmutationsaswellascommonvariantsfro

6、mwell-characterizedsamplesareidentified.Single-moleculesequencinggeneratesveryreproduciblecoveragepatternsandthesecanbeusedtodetectanysizeinsertionordeletiondirectly,unlikePCR-basedmethodswhichrequireadditionalassays.Becausenogeneisolationoramplificationisrequiredforsequencing,theexception

7、allylowcostsofsamplepreparationandanalysiscouldmakegenetictestsmoreaccessibletothosewhowishtoknowtheirowndiseasesusceptibility.Additionally,thisapproachhasapplicationsforsequencingintegrationsitesforgenetherapyvectors,transposons,retroviruses,andothermobileDNA