Molecular Biology Laboratory Manual

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1、MolecularBiologyLaboratoryManualP.Oelkers(June,2016version)TableofContentspg.WelcometotheMolecularBiologyLaboratory1OverviewofTranscriptionalRegulationProject:Promoter-ReportConstructStrategy3OutlineofTranscriptionalRegulationProject:Promoter-ReportCons

2、tructStrategy7AgaroseGelElectrophoresis11β-galactosidaseassays17BacterialTransformation20DesigningaScientificProject23DNAExtractionfromAgaroseGel27LaboratoryNotesandSettingUpaGoogleSite29Ligation32Microcentrifuge34Micropipeting35Oligonucleotide(e.g.prim

3、ers,oligos)DesignforPCR37OligonucleotideHandling43PolymeraseChainReaction45RestrictionEnzymeDigestion51ScientificLiteratureSearching55Yeast(S.cerevisiae)Culturing58Yeast(S.cerevisiae)GenomicDNAIsolation61Yeast(S.cerevisiae)RNAExtraction64Yeast(S.cerevis

4、iae)Transformation67WelcometotheMolecularBiologyLaboratorySciencecanbedefinedinavarietyofways.Sciencemaybetheconstructionofrulesandrelationshipsthatdefineaworldthatmatchesourworldascloselyaspossible.Sciencemaytheacquisitionofknowledgethroughinformationc

5、ollectionandanalysis.Inanycase,scienceinvolveslearning.Itinvolvestakingwhatisknownandexpandingandrefiningthatknowledge.Whilescienceitrootedinworkalreadydone,itsbranchesstretchoutintoaseeminglyendlesssky.Scientistsareexplorerswhogobeyondwhatitiscurrently

6、known.Withtheseloftyidealsinmind,eachgroupoftwoorthreestudentswillbeconductinganindependentresearchprojectduringthecourseoftheterm.Intheeducationfield,thisiscalledexperientiallearningorinquiry-basedlearning.Aframeworkfortheprojectswillbesuggested.Thisfr

7、ameworkwaschosensothatavarietyoftechniquesareemployed,thelogisticsarefeasible,andplanningcanbecompletedintwoorthreeweeks.Projectsinvolvingmolecularbiologyoutsidetheproscribedframeworkarepossible.Pleasediscusssuchprojectswiththeinstructorpriortoinitiatio

8、n.Asinanyproject,agoal-centeredapproachandattitudewillberequired.Toputitsimply,getitdone,well.Ifacertainexperimentalstepinunsuccessfulinachievingtheexperimentalgoal,thatstepwillhavetoberepeatedoranalternatestrategyadopted.Progres