Next-Generation Phylogeography A Targeted Approach for Multilocus Sequencing of Non-Model Organisms 英文参考文献.docVIP

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Next-Generation Phylogeography A Targeted Approach for Multilocus Sequencing of Non-Model Organisms 英文参考文献.doc

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Next-Generation Phylogeography A Targeted Approach for Multilocus Sequencing of Non-Model Organisms 英文参考文献

Next-GenerationPhylogeography:ATargetedApproach forMultilocusSequencingofNon-ModelOrganisms JonathanB.Puritz1*,JasonA.Addison2,RobertJ.Toonen1 1Hawai’iInstituteofMarineBiology,UniversityofHawai’iatMaˉnoa, Kaˉne’ohe,Hawai’i,UnitedStatesofAmerica,2DepartmentofBiology,UniversityofNewBrunswick, Fredericton,NewBrunswick,Canada Abstract ThefieldofphylogeographyhaslongsincerealizedtheneedandutilityofincorporatingnuclearDNA(nDNA)sequences intoanalyses.However,theuseofnDNAsequencedata,atthepopulationlevel,hasbeenhinderedbytechnicallaboratory difficulty,sequencingcosts,andproblematicanalyticalmethodsdealingwithgenotypicsequencedata,especiallyinnon- modelorganisms.Here,wepresentamethodutilizingthe454GS-FLXTitaniumpyrosequencingplatformwiththecapacity tosimultaneouslysequencetwospeciesofseastar(MeridiastracalcarandParvulastraexigua)atfivedifferentnDNAloci across16differentpopulationsof20individualseachperspecies.Wecompareresultsfrom3populationswithtraditional Sangersequencingbasedmethods,anddemonstratethatthisnext-generationsequencingplatformismoretimeandcost effectiveandmoresensitivetorarevariantsthanSangerbasedsequencing.Acrucialadvantageisthatthehighcoverageof clonallyamplifiedsequencessimplifieshaplotypedetermination,eveninhighlypolymorphicspecies.Thistargetednext- generation approach can greatly increase the use of nDNA sequence loci in phylogeographic and population genetic studiesbymitigatingmanyofthetime,cost,andanalyticalissuesassociatedwithhighlypolymorphic,diploidsequence markers. Citation: Puritz JB, Addison JA, Toonen RJ (2012) Next-Generation Phylogeography: A Targeted Approach for Multilocus Sequencing of Non-Model Organisms.PLoSONE7(3):e34241.doi:10.1371/journal.pone.0034241 Editor:BengtHansson,LundUniversity,Sweden ReceivedJanuary25,2012;AcceptedFebruary24,2012;PublishedMarch28,2012 Copyright: ? 2012 Puritz et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribut