Simultaneous Detection of FISH Signals and Bromo-Deoxyuridine Incorporation in Fixed Tissue Cultured Cells 英文参考文献.docVIP

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Simultaneous Detection of FISH Signals and Bromo-Deoxyuridine Incorporation in Fixed Tissue Cultured Cells 英文参考文献.doc

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Simultaneous Detection of FISH Signals and Bromo-Deoxyuridine Incorporation in Fixed Tissue Cultured Cells 英文参考文献

SimultaneousDetectionofFISHSignalsandBromo- DeoxyuridineIncorporationinFixedTissueCultured Cells DanielaMoralli,ZoiaL.Monaco* WellcomeTrustCentreforHumanGenetics,UniversityofOxford,Oxford,UnitedKingdom Abstract FISH (Fluorescence in situ hybridization) is a powerful technique that detects and localises specific DNA sequences on metaphasechromosomes,interphasenucleiorchromatinfibres.WhencoupledtoBrdU(5-Bromo2-deoxy-uridine)labeling ofnewlyreplicatedDNA,thereplicationpropertiesofdifferentDNAsequencescanbeanalysed.However,thetechniquefor thedetectionofBrdUincorporationistimeconsuming,andreliesonacidicpHbuffertreatments,thatpreventuseofpH sensitivefluorochromessuchasFITC(Fluoro-isothiocianate)duringFISH.Inthiswork,wedescribeasimplifiedprotocolthat allows the simultaneous detection of FISH signals and BrdU incorporation. Since the technique does not involve paraformaldehydeforcellfixation,orformamidefordenaturationofthetargetDNAandinpost-hybridisationwashes,it representsasaferalternativetoclassicalFISHtechniques. Citation: Moralli D, Monaco ZL (2009) Simultaneous Detection of FISH Signals and Bromo-Deoxyuridine Incorporation in Fixed Tissue Cultured Cells. PLoS ONE4(2):e4483.doi:10.1371/journal.pone.0004483 Editor:JoannaMaryBridger,BrunelUniversity,UnitedKingdom ReceivedDecember4,2008;AcceptedJanuary6,2009;PublishedFebruary16,2009 Copyright: ? 2009 Moralli et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwasfundedinpartbytheWellcomeTrustandtheDystrophicEpidermolysisBullosaResearchAssociationUK.Thefundershadnoroleinthe studydesign,datacollectionandanalysis,decisiontopublish,orpreparationofthemanuscript. CompetingInterests:Theauthorshavedeclaredthatnocompetinginterestsexist. *E-mail:zoia@well.ox.ac.uk Introduction signalscanbevisualisedusingspecificantibodies.However,dueto thepos