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1、NIHPublicAccessAuthorManuscriptScience.Authormanuscript;availableinPMC2013June21.NIH-PAAuthorManuscriptPublishedinfinaleditedformas:NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptScience.2012December21;338(6114):1622±1626.doi:10.1126/science.1229164.Genome-WideDetec
2、tionofSingleNucleotideandCopyNumberVariationsofaSingleHumanCellChenghangZong1,?,SijiaLu1,?,#,AlecR.Chapman1,2,?,andX.SunneyXie1,*1DepartmentofChemistryandChemicalBiology,HarvardUniversity,Cambridge,MA02138,USA2PrograminBiophysics,HarvardUniversity,Cambridge,MA021
3、38,USAAbstractKindredcellscanhavedifferentgenomesbecauseofdynamicchangesinDNA.Singlecellsequencingisneededtocharacterizethesegenomicdifferencesbuthasbeenhinderedbywhole-genomeamplificationbias,resultinginlowgenomecoverage.Herewereportanewamplificationmethod:Multi
4、pleAnnealingandLoopingBasedAmplificationCycles(MALBAC)thatofferhighuniformityacrossthegenome.SequencingMALBACamplifiedDNAachieves93%genomecoverage≥1xforasinglehumancellat25xmeansequencingdepth.Wedetecteddigitizedcopynumbervariations(CNVs)ofasinglecancercell.Byseq
5、uencingthreekindredcells,wewereabletocallindividualsinglenucleotidevariations(SNVs)withnofalsepositivesobserved.Wedirectlymeasuredthegenome-widemutationrateofacancercelllineandfoundthatpurine-pyrimidineexchangesoccurredunusuallyfrequentlyamongthenewlyacquiredSNVs
6、.Singlemoleculeandsinglecellstudiesrevealbehaviorsthatarehiddeninbulkmeasurements(1,2).Inahumancell,thegeneticinformationisencodedin46chromosomes.Thevariationsoccurringinthesechromosomes,suchassinglenucleotidevariations(SNVs)andcopynumbervariations(CNVs)(3),areth
7、edrivingforcesinbiologicalprocessessuchasevolutionandcancer.Suchdynamicvariationsarereflectedinthegenomicheterogeneityamongapopulationofcells,whichdemandscharacterizationofgenomesatthesinglecelllevel(4±6).Singlecellgenomicsanalysisisalsonecessarywhenthenumberofce
8、llsavailableislimitedtofeworone,suchasprenataltestingsamples(7,8),circulatingtumorcells(9),andforensicspecimens(10).Promptedbyrapidprogressinnextgenerationsequ
