第1題IntheprocessofgenomicDNAextraction,thepossibleoperationsthatcausethedegradationandfragmentationofmacromolecularDNAareAEquipmentsterilizationtreatmentBGentleoperationCAddEDTADRepeatedlypipettingtheDNApellettopromotedissolution第2題TheroleofEDTAinthecelllysisbufferusedtoextractgenomicDNAfromtissuesorcellsisADestroycellmembraneandnuclearmembraneBSeparatetissueproteinfromDNACInhibittheactivityofDNaseDDegradationofprotein第3題TheroleofproteinaseKusedtoextractgenomicDNAfromtissuesorcellsisADestroycellmembraneandnuclearmembraneBSeparatetissueproteinfromDNACInhibittheactivityofDNaseDDegradationofprotein第4題TheroleofRNaseAusedtoextractgenomicDNAfromtissuesorcellsisADestroycellmembraneandnuclearmembraneBSeparatetissueproteinfromDNACInhibittheactivityofRNaseDDegradationofprotein第5題TheratioofpureDNAOD260/OD280isaboutA1.8B1.6C1.5D2第6題AfterDNAextraction,iftheratioofOD260/CD280is2.0,thepossiblereasonisASaltorsmallmoleculepollutionBRNAcontaminationCProteincontaminationDOrganicsolventcontamination第7題AfterDNAextraction,iftheratioofOD260/CD280is1.5,thepossiblereasonisADNAdegradationBRNAcontaminationCProteincontaminationDSaltpollution第1題TheplasmidusedinDNArecombinationtechologyis：APartofthecellchromosomalDNABtheindependenthereditaryunitoutsideofthebecterialgenomicDNACpartoftheviralgenomicDNADpartoftheeukaryoticchromosomegenomicDNA第2題restrictionendonucleaserecognizethespecialstructureofDNA,thespecialstructureiscalled()ApalindromicstructureBmotifCsupersecondarystructureDtertiarystructure第3題TheenzymecanrecognizespecialsequencesandcleaveDNAatthesespecificbasesequencesiscalled?ADnaseBRestrictionextronucleaseCRestrictionendonucleaseDnon-Restrictionendonuclease第4題WhatarethemaincomponentofLysisBuffer?Asodiumhydroxide(NaOH)andSDS.BglucoseandTris-HClCEDTAandRNaseDAcetateandPotassiumacetate第5題WhichkindofelectrophoresisisfrequentlyusedtoanalyzeplasmidDNAasapowerfulseparationmethod?ASDSBAgrosegelelectrophoresisCPCRDWestern-BlotPrinciple,extractionoperationandidentificationofpolymerasechainreaction第1題PCRcomponentsinclude()AprimersBtemplateCTaqpolymeraseDNTP正確答案：ABC第2題PCRprocessincludesthreesteps,theyare()AdenaturationBannealingCextentionDcooling正確答案：ABC第3題Simplesetofrulesforprimersequencedesigninclude()APCRprimersshouldbe15-30nucleotidesinlength.BoptimalGCcontentoftheprimeris40%-60%.CpreferoneortwoGorCatthe3’-endofprimer.Davoidprimerselfcomplementarities.正確答案：ABCD第4題WhataretherequirementsforDNAtemplateinPCR?()ApurifiedDNAtemplate.Buse1pg-1ngofplasmidorviraltemplate.Cuse1ng-1μgofgenomictemplates.DUsethehigherDNAconcentrationswhenfewercycledesired.正確答案：ABCD第5題WhenweamplifyatargetDNAsequencebyPCR,howtodetermineannealingtemperature?()ATmBTm-5℃C55℃D60℃Principle,operationandcalculationofLowrymethodforproteindetermination第1題Thetotalproteinconcentrationisexhibitedbyacolorchangeofthesamplesolutioninproportiontoproteinconcentration,whichcanthenbemeasuredusing().AbalanceBspectrophotometerCcentrifugeDpipette第2題TheLowrymethodisbasedonthereactionof（）producedbytheoxidationofpeptidebondswithFolinreagentunderalkalineconditions.ACu2+BFe3+CMg2+DCa2+第3題ThereactionsinLowrymethodresultinastrongbluecolor,whichdependsmainlyontheaminoacidsresidues,suchas()and(),theintensityofwhichisrelatedtothecontentofprotein.AAlaGlyBArgAlaCTrpTyrDGluGly第4題InLowrymethod,thesamplesandthestandardproteinsolutionaretreatedidentically.Aftercolordevelopment,theabsorbanceofeachsampleisdeterminedbyabsorbanceat()nm,thustheproteincontentcanbecalculated.A550B710C280D650第5題ThecompositionofFolinreagentisamixtureof().AphosphomolybdateandphosphotungstateBHClandphosphomolybdateCphosphotungstateandH3PO4DphosphomolybdateandH3PO4Principle,operationandresultdisposalofSDS第1題WhichisrightfortheSDSPAGE?AintroductionofSDSsysteminpolyacrylamidegelBthemigrationdistanceisrelatewithproteinmolecularandchargeCtheconcentratedgelbufferispH10.0DtheseparationgelbufferispH10.0第2題Whenthemolecularweightfrom15kDto200kD,whathavelinearrelationshipwiththelogarithmofthemigrationrate?AmolecularweightofproteinBtheshapeofproteinCthechargeofproteinDthepIofprotein第3題Insaturatedstate,howmanySDSbindwithpergramofpeptide?Aabout10.4gramBabout0.4gramCabout1.4gramDabout0.14gram第4題InSDSPAGE,thegelbufferusedis?ATris-GlycineBTris-HClCNa2CO3-NaHCO3K2HPO3-KH2PO3第5題whatisthefunctionofbetamercaptoethanolwhenyoudealwiththesample?AbreakthehydrogenbondBbreakthe-S-S-bondCbreakthepeptidebondDbreakthephosphatebondPrinciple,operationandresultofAffinitychromatography第1題ProteinsseparationcanbecarriedoutonthebasisofAnetchargeBsolubilityinsaltsolutionsCsizeormassDallofthese第2題AffinitychromatographydealswiththeAspecificbindingofaproteinconstituentsforanothermoleculeBprotein-proteininteractionCprotein-carbohydrateinteractionDcarbohydrate-carbohydrateinteraction第3題Therearemanytypesofchromatography.ThereareAaffinitychromatographyBgelfiltrationchromatographyCion-exchangechromatographyDallofthese正確答案：第4題TheadvantageofaffinitychromatographyisAmoreefficientBverycheapCcanseparateallmacromolecularDcanseparateallmacromolecular第5題Theappropriateligandisattachedtothematrixofthecolumnandaproteinextractisrunthroughthecolumn.Theonlymacromoleculethatwillattachtothecolumnwillbetheonethatbindstotheligand;alltheothermacromoleculeswillwashthroughthecolumn.Thecolumnisthentreatedinsuchamannerthattheligandandthemacromoleculewilldissociateandthemacromoleculewilltheneluteoff.Elutionbyinclusionoffreeligandinbufferiscommon.OthermethodsofelutioncanbeAloweringthepHofbufferBincreasingordecreasingsaltconcentrationsCdecreasingthetemperatureofthecolumnDallofthese正確答案：Principle,operationandcalculationofKmvalueofalkalinephosphatase第1題WhichoneistheMichaelis-Mentenequation?()ABCD第2題TheMichaelis-Mentenequationdemonstratesthatwhenv=?Vmax,whichoneisthesame