1、甘肅農(nóng)業(yè)大學(xué)碩士學(xué)位論文PCR技術(shù)在羊早期胚胎性別鑒定中的應(yīng)用姓名：李小英申請學(xué)位級別：碩士專業(yè)：動物遺傳育種與繁殖指導(dǎo)教師：崔峴劉月琴20050601SummaryT、voDaisofnestedPCRprimersweredesignedtoidentifytheearlyembryossexofsheepandgoataccordingtothehomologyinbovine，sheepandgoatSRYgeneMeanwhi

2、le，meotherpairofprimerwerealsodoneinvirtuetosequencesofsheepp—globingeneasinternalcontrolgeneprimerAccordingtothiscombination，aPCRsystemofidentifyingsheepandgoatembryoswasestablishedAstonestedPCR，twotempreturecirculation

3、amplificationwascomparedwiththreetempreturecirculationaboutamplifiedtimeandresultsGenomicDNAfromhuman，mouseandcattlewereamplifiedtoassaythespecificityofSRYgeneprimerandinternalcontrolgeneprimerInadditionto，thestudywasper

4、formedonwhethersomepossiblefactorscontaminatethesampleinsexdeterminationofsheepandgoatembryosAtotalof44malegenomicDNAsamplesfromthebloodincludingtosheepandgoatsand19femaleSeXwasindentifiedbydetectingSRYgenewithnestedPCRp

5、rocedureThemalesamplehad170bpfragmentofamplifiedSRYgeneand240bpfragmentofamplifiedinternalcontrolgene；thefemaleonlyhad240bpfragmentbandThepredeterminedsexeswerefullyinaccordancewiththeirrealsexes，theaccurateratewas100％It

6、wasproventhattwo—tempreturecirculationPCRcouldefficientlydecreasethetimeofamplificationopposedtothreetempretureciculationamplification，furthermore，theresultswerealmostsameBytestingthespecificityofprimers，SRYgeneprimerwas

7、specifictocattle，sheepandgoatTheoperators’sexhadnotinfluencedontheresultofidentification44sheepembryosand26goatsembryosfromsuperovulatedsheepandgoatswereselectedtoindentifywithestablishedPCRsystemTheamplificationresultwa

8、sdeterminedsexesofembryosefficientlybyassayingthesexof1／4’115embryoThesplittingwasperformedusingabio—cutbladeinhandandNARISHIGEmicromanipulatortoinvestigatetheeffectofdifferentmethodsonthepracticeduringsplittingmouseembr

9、yosatmorulaeandblastocystsThesectioninhandwasmoresensitiveusefulinspiteoftheunexactThesheepsertlmincollectingmedumandholdingmedumcannotcontaminatethesampleandidentilyingresultsofearlyembryosThesystemwaspracticalKeywords：