海帶孢子體DNA的提取及其多態(tài)性研究.pdf

1、上海水產(chǎn)大學(xué)碩士學(xué)位論文海帶孢子體DNA的提取及其多態(tài)性研究姓名：胡遠(yuǎn)皆申請學(xué)位級別：碩士專業(yè)：水產(chǎn)養(yǎng)殖指導(dǎo)教師：周志剛2001.6.1塑堊堂圭塑查蘭奎蘭望主蘭垡堡莖!!!!蘭!旦圭塑!!!!!!ExtractionandPolymorphismofDNAinLaminariajaponica(Phaeophyta)Sporophyte(ExperimentalBiotechnologyofAlgae，CollegeofFisheryS

2、cience，KeyLaboratoryofEcologyandPhysiologyinAquaculture，theMinistryofAgricultureShanghaiFisheriesUniversity，334JungongRoad，Shanghai200090，PRChina)AbstractAnewextractionprotocoltoobtainhighqualityDNA，whichinvolvestheenzym

3、aticdissociationofLaminariajaponicasporophytetissuesandtheeliminationofremainingpolysaccharideswithcetyltrimethylammoniumbromide，hasbeendevelopedUnicellsisolatedfromthefrozenkelptissueswithalginatelyasepreparedfromtheaba

4、loneHaliotisdiversicolorwereusedtoextracttotalDNAandtheyieldwasapproximately13to225gDNAg。(wetsporophyteweight)TheaveragesizeofgenomicDNAwasaround23kbestimatedbyagarosegelelectrophoresis，andthepurityoftotalDNAdeterminedsp

5、ectrophotometricallyastheratioofOD2clOD280wasabout17TheingredientsandprogramsofpolymeraseamplifiedreactionswereindividuatlyoptimizedThereactionswerecomposedof02uMprimer，15mMM92100uMdNTP，10UTaqDNApolymerase，30ngtemplateDN

6、ATheoptimizedprogramswerepredenaturationat94“Cfor2min；andthen45cyclescontainingdenaturationIrainat94。C，annealing1minat37℃，andelongation2minat72“C；andatlastholding10minat72“CTheoptimizedconditionscouldresultinsatisfactory

7、specificityandreproducibilityofRAPDamplificationpatternsRandomamplifiedpolymorphicDNA(RAPt))markerswereusedtodetectsixdifferentcultivarsLaminaria、Ⅳim30primersAtotalof185locusweredetectedand76ofallthebandsshowedthepolymor