Isolation and characterization of polymorphic microsatellite loci

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1、ConservationGenetResourDOI10.1007/s12686-014-0263-zMICROSATELLITELETTERSIsolationandcharacterizationofpolymorphicmicrosatellitelocifromSilurusLanzhouensisYuan-HaoYang•FengWang•Lv-ZhouWang•LeiLi•Juan-NingYang•YangLiReceived:8June2014/Accepted:7July2014ÓSpringerScience+BusinessMedia

2、Dordrecht2014AbstractSilurusLanzhouensisisanendangeredfresh-toresurrectthestocksofthespeciesthroughconservationwaterfishendemictoNingxia,GansuandShaanxiprov-andfisherymanagement,butthesystematicpositionandinceofChina.Herewedescribeafirstsetof12populationstructureofthefishislargelyunkn

3、own.polymorphicSilurusLanzhouensismicrosatelliteloci.TheMicrosatelliteDNAmarkersseemtobeusefulfortheobservednumbersofallelesrangedfrom2to7.Thegeneticassessmentsofthefish,buttillnowthereisnoexpectedandobservedheterozygositiesrangedfromspecies-specificmicrosatellitemarkerhasbeenreport

4、ed.In0.3279to0.8438andfrom0.4021to0.8410,respectively.thepresentstudy,weisolatedthefirstsetof12informativeThesemicrosatellitemarkersareusefulforpopulationandmicrosatellitelociusingfastisolationbyAFLPofconservationgeneticstudiesinS.Lanzhouensis.sequencescontainingrepeats(FIASCO)prot

5、ocol(Zaneetal.2002)withslightmodifications.KeywordsSilurusLanzhouensis
Threatenedspecies
TotalgenomicDNAwasextractedfromthefishfinsFIASCO
MicrosatellitesusingatraditionalproteinaseKandphenol–chloroformmethod.Atotalof250nggenomicDNAfromapoolof40SilurusLanzhouensis,orYellowRivercatfish,

6、isalarge-specimenswascompletelydigestedwithMseI(BioLabs).sizecarnivorousfishbelongingtothefamilyofSiluridea.ItThedigestedDNAfragmentsofbetween400and1,200bpisendemictotheupperreachesoftheYellowRiverinwererecycledbyaDNAGelRecycleKit(Sangon)andthenligatedtoMseIAFLPadaptor(50-GACGATGAG

7、Ningxia,GansuandShaanxiprovinceofChina(Wuetal.TCCTGAG-30/50-TACTCAGGACTCAT-30)withT4DNA2006).Duetoenvironmentaldegradationandillegaloverfishing,thepopulationsizeofthefishdeclinedsig-ligase(BioLabs).Theligationmixwasdilutedto1/10,andnificantlyinrecentyears.Consequently,S.Lanzhouensisd

8、irectlyamplifiedusingMseI-Nadaptor