應(yīng)用微衛(wèi)星標(biāo)記對(duì)雞屠體性狀和生長性狀的初步研究.pdf

1、江西農(nóng)業(yè)大學(xué)碩士學(xué)位論文應(yīng)用微衛(wèi)星標(biāo)記對(duì)雞屠體性狀和生長性狀的初步研究姓名：黃銀花申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：動(dòng)物遺傳育種與繁殖指導(dǎo)教師：孫漢李寧2001.6.1皇里絲墨呈堡里型墮星堡絲鑒塑生量絲鑒塑塑生塹塞AbstractGroatprogresshasbeenachievedinpoultrybreedinginlastcenturyThemajorlimitationofthepreviousstudywasthequantitati

2、vetraitlocuscouldnotbemappedandclonedWhⅡethenewadvancessuchasgenomicscanningthroughmierosatellitehavemadeitpossibleforquantitativetraitlocusanalysis，gonecloningandfunctionanalysisIn370F2chickensofCAUresourcefamilyderived

3、frombroilerscrossingtosilk)；therelationsbetweenthegenesandcarcassaswellasgrowthtraitshadbeendetectedthroughmicrosatellitebymutiplexPCRandfluorescencebasedsemiautomatedelectrophoresisInorder幻takeadvantageofthemuffplexPCRi

4、t’Sconditionshadbeenoptimizedindividuallyandincombinationincludingtheannealingtemperatures(4255“C)，thelengthofthepolymerizationstep(30s1rain)，thefinalelongationste“2040min)，thecyclenumbersinthecyclingprogram(eachannealin

5、gtemperaturecychng10～20)，thecontentofthePCRbufferandthevolumeofthereactionmix(1525u1)HEX，TETor6FAMamiditefluorescenceWaslabeledatthe57or37endoftheprimersAccordingtothefluorescentdifference，fragmentlengthandprimersparinge

6、liminationPCRworeperformedwithbothasinglemarkerPCRandmudplexPCRamplificationforalIF0individualsOnlythemarkerswiththeidenticalamplifiedfragmentsinbothPCRreactionsweroselectedforgeneticlinkageandquantitativetraitslocationa

7、nalysiswithinFlandF2populationsnleoutputsofmutiplexPCRweredilutedandscreenedintheGS36C2400modulefor3hOBABI377SequencerDNAfragmentanalysiswereperformedusingGeneScan3landGenotyper25(PcrkinEImor)underthemanufacture’SmanualP

8、eaksizeswerecalculatedusingthe‘‘LocalSouthernMethod’sizingoptionOnthebasisofthisoptimizationexperiments170outof328polymorphicmicrosatellitemakerswereclassifiedinto30differentsetsTheycouldbeappliedingenotypingviatllemutip

9、lexPCRinthemappingofquantitativetraitlocusTheoptimizationtestresultsindicatedthatthecyclingparametersworethecriticalfactorsf＆memutiplexPCRreactions，ietheannealingtemperatures，theannealingandelongatingtime，andthefinalelon

10、gatingtime。OtherinfluencingfactorswerethePCRbuffercontentandDNAqualityWhilethemutiplexPCRreactionvolumeisnotsignificantDataanalysiswasperformedwithSAS61TherasultsindicatedthatcarcassandgrowthtraitsassociationwithLEl85，LE