【生物科学】分子细胞学技术

12DNADNARecombinationDNADNAGeneEngeneeringDNADNAGeneticEngeeringDNA80318652050CrickWatsonDNADNA4DNARNARNADNARNARNADNADNADNADNA461.6m~8.2m46DNAGenomeDNA3109bp5~10DNADeoxynucleotideDNADNAAGCTURNAdNTPdNTPdATPAdTTPTdCTPCdGTPGDNA123451234553355Upstream3Downstream53DNADNADNAA-TC-GDNA13-1DNADNAA-TG-C513-1DNA6DNACodeDNARepeatedSequenceCopy2~10AlurRNAtRNA1~3kbBasePairbprRNAtRNARegulaterPro-moterEnhencerSilencerrRNAtRNARegulaterPro-moterEnhencerSilencer2~305353Poly-morhism463109bp50000~100000130Mb106bp2000~4000DNADNADenaturation7DNAG-CA-TA-TG-CG-CA-TDNADNADNADNADNADNADNADNAS13-2DNADNADNAO.D.Opti-calDensityAnnealDNADNADNADNADNADNADNADNADNADNADNADNA30DNADNAE.Coli20DNAE.ColiDNA403DNA13-3DNADNAPrimerRNADNARNA3DNADNAIII53A-TG-C105bp5102~5103bp1ExpressionCodeDNARNADNARNATran-scriptionRNATranslationDNARNADNASenseStrandPositiiveStrandAntisenseStrandNegativeStrandDNADNAmRNAhnRNA813-2DNA9SplicingRNAmRNA13-3mRNAGenotypePhenotype13-310Jacob-MondRNARNACis-actingElementsPromotorEnhancerSilencerDehancer5`100bpGoldberg-HognessTATATATAPrinbnowTATA30bpTATATATATrans-ActingElementsDNADNADNABindingProteinDNA13-4RNADNARNADNAU10-5gRNA5~10gRNA1108RNARNA13-1RNA%rRNARNA80%~85%TrnaRNARNA10%~15%MrnaRNA1%~5%mRNAmRNAmRNAmRNA320~250PolyAdTRNARNaseRNARNase1113-4DNADNASomaticCellDNAGermCellGenomeAlleleLoeusDominantRecessiveHomozygoteHeterozygoteDNADNADNA12PointMutationDNARearrage-mentAberrationDNADNADNADNADNADNADNADNADNADNA13,EthydiumBromideDNADNAAmesMethyleneBlueDNA,5mg/ml1l/ml,a0.5mg/mlb10.5MKMnO4KMnO4c12.5NHCIde12.5NNaOHfDNANIHE.ColiK1219221970DNA14CO20.22m1.5mlEppendorf15ml25ml15pHGTL-16G15000/0~500V0~150mA400~1000V1000VDNADNADNA3PharmaciaRNA4DNADNAATCG5ddNTPPCRPolymeraseChainReactionPCRDNADNADNADNAPCR16XChromatographyKdKd=CmCsKdCmCs1714-1SephadexSepharoseAllyldextranNN-SephacrylBiogel-PSephadesSephadexSphadexG-501500~30000[-32P]dNTPSepharoseAbiogel-ApH4~9402B4B6B2%4%6%SepharoseSephadexDNASephacrylpH3~11DNAdNTPSephadex1SephadexG-50DNADNAdNTPDNA3[-32P]dNTPcDNADNA1814-1SephadexG-50DNADNA100~200lTEpH7.6SephadexSephadexSephadex1910gSephadexG-50250ml500ml651~2DNATEpH7.6415155mlSepdaesx5ml1102ml1Sephadex25TE1ml/3DNA200lDNA100lTEpH7.65TETETE0.3ml/10~15630%1~37TETE8cpmcpm-20,0.5~1.0gDNAcpm3300~400lDNA3Sepharose4BpolyAmRNAdtUD-dTA-U20polyARNAmRNAmRNADNAHgRNA-Hg-NTP2-SH-NTPSHHgSepharoseRNAIonExchangeChromatographyIECIECpI2~2.5[DEAEC2H52+NH-CH2CH2O-]tRNArRNANaCltRNArRNAHydroxyapatideHACa5PO43OHHAHAcDNAHAReversedPhaseChromatographyRPCRPC-1RPC-2RPC-5RPCtRNAtRNA10~130merpolyA+1DNA4DNAGCATDNARPC70~4000bpDNACCDNACCDNA1/321DNARNAK/RNAase10%~15%DNApolyARNA6810121NaCl2DNARNADNARNADNADNADNADNADNASDSKSDSEDTADNASDS22EDTADNaseKDNARNaseRNA/DNADNADNASouthern3123156PBS10mMTrispH7.410MEDTA150MmNaCl0.4%SDSK20mg/ml50~100g/mlTrisSS-TERNaseADNase10mg/ml10mg/mlRNase10mMTrisHCIpH7.5100103MpH7.4TE1106~109DNAPBSTBS22K10K365154K37511/30ml12000g106756830ml1/103M2.5DNA912000g10DNA80%5ml23104.5mlTEDNA37RNaseARNA11DNATERNaseADNase20g/ml373012500l3M5ml11/1312000g10145ml131512.5ml2.5DNA1612000g5m180%1718TEDNA3719OD260mDNADNADNAPBSKDNADNA50%DNADNA37DNADNA1mg/mlDNAKDNaseRNaseARNaseTE10mg/ml10010DNaseDNADNADNADNADNA100~150kbSouthernDNADNA414-280%DNA100kbDNA12414-2DNA210K350ml3717DNA[3]DNADNADNA18~20mlAcidCitricDetroseSolutionACD2%EDTA2gEDTA+0.85gNaCI+H2O100mlACDACD0.48g1.32g1.47g100.0mlACD6ml1mlACDACDEDTAACD470220ml1300g15PBS35001515ml371315ml3714K100g/mlDNA[3]25RNAmRNAmRNAmRNARNARNADNARNARNARNAmRNAhnRMARNArRNAtRNAmRNARNaseRNaseRNARNARNaesRNARNaseRNaseRNaseRNA1RNase0.1%DiethylPyrocarbonateDEPCDE-PC1604EppendorfRNaes0.05~0.1%DEPCDEPCDEPCcDNAPCRDEPCRNaseSDSBentoniteRNaseRNARNAKRNaesRNaseRNARNaesRNARNaes1.78g/mlCsClRNADNARNADNABeckmanSW41SW550.3MpH6.03.0MpH6.0tRNA10mg/mlGIT94.53gGIT4M1.67ml3.0MpH6.0200ml0.8m261.67ml--95.97g5.7M0.83ml3MpH6.0100ml0.8m80%1RNA80%RNA24106GIT4mlGIT310ml4mlCsClCsCl4GITSW4132000rpm174000g20215200lRNA6200l0.3MpH6.0RNAEppenedorf100l0.3M7300l18750l10910300l80%4250~100l10OD260nmRNAK/RNAcDNASIRNA30~500g/107RNA0.14MNaCl1.5mMMgCl210mMTrisClpH8.60.5%NP-401mM2-1000U/mlRNaseI20mM27KTrisClpH8.00.2MEDTApH8.025mMNaCl0.3MSDS2%K20mg/mlRNaseDNaseITE1a7mlPBS1bPBSEppendorfc2000g4510PBS2d1mlPBSEppenddorf212000g430200lRNA155312000g41.514200lKK50g/ml305/5000g1061Eppendorf40l30712000g41070%RNA8200lTEpH7.6RNA500l70RNADNADNARNADNARNAcDNANorthern97RNA20l50mMTrisClpH7.81mMEDTApH8.010MgCl22-10mM0.1mMRNaes1000U/ml10mM11RNaseDNaseI2g/ml37602812EDTASDS10mM0.2%13/12000g5Eppedorf1412000g45200lTERNA500l70mRMA5m7G3polyAmRNA320~300polyApolyA+mRNAdTdUmRNAmRNAdT-NaOH0.1M1Tris.ClpH7.620mMNaCl0.5MEDTApH8.01mMSLS0.1%Tris.ClpH7.6NaClEDTApH8.065653010%SLS0.1%Tris.ClDEPC10mMTris.ClpH7.61mMEDTApH8.00.05%SDSTris.ClEDTA10%~20%SDS3MNaAcpH5.210.5~1.0gdT-0.1MNaOH20.1%DEPC1mldT-0.1~1ml331pH8.04RNA6552RNA291565565~1011mlOD269RNAOD260polyARNAOD2600.1MNaCl51polyARNA72~3polyARNA1/3~1/28OD260polyARNApolyARNA1/103MNaAcpH5.22.5700.13MNaAcpH5.212000g45polyARNARNase4RNA655RNAmRNApolyA+polyA+polyA+RNAmR-NArRNArRNA7NaCl0.5M6555-7polyA+RNAdT4NaOH1OD26040g/mlRNA1071~5gpolyARNARNA1%~2%DEPCDNARNA260nm280nmOD260nmOD150g/mlDNA40g/ml