【基础分子生物学】第5章.pdf
50页
2009/12/1122009/12/112 20 DNA PCR 2009/12/1132009/12/113 2009/12/114 1.DNA 2. DNA 3. RNA 4. 5. 2009/12/114 2009/12/115 DNADNA 2009/12/115 40 DNA 50DNA , 5060“” , 2009/12/1162009/12/116 DNA 1869F MiescherDNA 1944O.T. AveryDNA 1952A.D. HersheyM.ChaseDNA 1953 J.D.WatsonF.H.C.CrickDNA M.WilkinsX- 1957A.KornbergDNAI 1958M. MeselsonF. W. StahlDNA 1959- 1960 S. OchoaRNARNAmRNA 1961 NirenbergF. JacobJ. Monod 1964 C. YanofskyS. Brenner 2009/12/1172009/12/117 1965 S. W. HolleytRNA DNA 1966 M.W.NirenbergS.OchoaH.G.KhoranaF.H.C.Crick 1970 H.O.SmithK.W.WilcoxT.J.Kelley H.M.TeminD.BaltimoreRNA 1972- 1973 H.BoyerP.BergDNA72 DNA73 1975- 1977 F.SangerA.MaxamW.GilbertDNA 19775387bp174 1978 1980 1981 R. D. PalmiterR. L. Brinster A. C. SpradlingG. M. Rubin 2009/12/1182009/12/118 1982- - Sanger48,502bp 1983 1984DNA 1986GMO 1988J. D. Watson 1989DuPont- - Oncomouse 199235 1994 1996(1.25107bp) 1997 2000 2001 2004 2005 2009/12/119 DNADNA DNADNA DNA DNA DNA EcoRIBoyer1972 GAATTCDNA 19675 DNA 2009/12/119 2009/12/1110 DNA 2009/12/1111 DNADNA a. DNAb. DNAc. EcoRIDNA 2009/12/1112 DNADNA DNA DNA DNA DNA DNADNA DNA 2009/12/1112 2009/12/1113 DNADNA DNA DNA3-5 DNA3 cDNA 5 3 3 DNA3 DNA5 DNA 2009/12/1114 DNADNA 1. DNA DNA pH 2009/12/1115 DNADNA 1. - DNARNA -DNA DNA DNA 2009/12/1116 DNADNA 1. DNA0.250kb 11000 EtBr DNADNA 0.05gDNA 2009/12/1117 DNADNA DNA DNA 2009/12/1118 DNADNA 50kbDNA 2009/12/1119 DNADNA 2.DNA DNA DNADNA 0 DNA 42 2009/12/1120 DNADNA 2.DNA CaCl2 DNA DNA DNA 2009/12/1121 DNADNA 3. PCR DNA DNA DNA DNADNADNA DNA DNA DNADNA() DNA()DNA() DNA DNA2n 2009/12/1122 DNADNA 3. DNA 941minDNA DNA 561min DNADNA 721DNA 3-53 DNAPCR PCR 2009/12/1123 DNA DNA 2n DNADNA 2009/12/1124 DNADNA 4.PCR PCRPCR PCR PCRPCR 2009/12/1125 DNADNA 4.PCR , PCR pcr Ct CtCt Ct 2009/12/1126 PCRpcr TaqManPCR PCRTaq 53 DNA PCR SYBRPCRSYBR SYBRDNA SYBR PCR 2009/12/1127 TaqManSYBR 2009/12/1128 DNADNA 4.DNA DNA DNA DNA DNADNA DNA DNA 2009/12/1129 DNA 2009/12/1130 DNADNA SNP Single nucleotide polymorphisms, SNP 1000SNP SNPSNP SNPsDNA 2009/12/1131 DNADNA SSR DNA 110bp TGTGTG= (TG)nAATAATAAT= (AAT)n SSR SSR SSR(Flanking Region) simple sequence repeats 2009/12/1132 RNARNA 1.RNA RNARNA LiClCTABTriZol TriZol TRIZOL RNA RNA RNA RNA DNARNA DNA 2009/12/1133 RNARNA 2. mRNA mRNAdT- mRNAPoly AdT Poly AdT- mRNA -SA- PMPs dT- mRNA- SA- PMPs SA- PMPs dT mRNAmRNA 2009/12/1134 PolyATtract mRNA 2009/12/1135 RNARNA 3. cDNA cDNAcDNA cDNAmRNAcDNA DNA oligo dTDNA 2009/12/1136 RNARNA 4. cDNA cDNAmRNA cDNA cDNAOligo(dT) cDNA mRNAcDNA DNAcDNA cDNAcDNA DNA12 DNA 2009/12/1137 RNARNA 5. DNA PCR 2009/12/1138 clone monozygotic twins progenitor cell DNA DNA 2009/12/1139 1. RACE cDNA PCR 55RACE 33RACE cDNAcDNA 2009/12/1140 1. RACE 5 RACE cDNA RNasemRNA cDNA cDNA3 dCTP oligo(dG) nested PCR nest PCR 2009/12/1141 1. RACE 3RACE polyAoligo(dT) cDNA RNaseHmRNA PCR3 nest PCR 2009/12/1142 2.cDNA cDNArepresentational difference analysis, RAD PCRDNA cDNA cDNA 2009/12/1143 3. Gateway Gateway Int integraseIHFIntegration Host Factor attBattPDNA53 attLattRIHF, IntXis E. coli DNA DNA53attLattR lambda DNA attPE. coli DNA attB 2009/12/1144 3. Gateway BPLR PCR PCR 2009/12/1145 4. RFLPRAPD RFLP 2009/12/1146 2009/12/1147 1. 2- DEOFarrells1975 1 000E.coli pH Internet 2- DE 2009/12/1148 1. pH3 pH10IEF SDS 2009/12/1149 2. Western blotingimmunobloting PAGE 2009/12/1150 2. SDS- PAGE 2009/12/1151 2. 3. 。
