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with PCRE_UTF8 support at offset 0 in /web/Sites/BlickinsBuch.de/functions.php on line 241 Warning: preg_replace(): Compilation failed: this version of PCRE is not compiled with PCRE_UTF8 support at offset 0 in /web/Sites/BlickinsBuch.de/functions.php on line 241 Blickinsbuch.de - Molekularbiologische Methoden - Thomas Reinard
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    Molekularbiologische Methoden

    Weitere Titel des Verlages

    Molekularbiologische Methoden

    Autoren:

    Verlag:
    UTB   Weitere Titel dieses Verlages anzeigen

    Erschienen: November 2010
    Seiten: 220
    Sprache: Deutsch
    Preis: 29.90 €
    Illustration: 127 Abbildungen, 41 Tabellen, 15 Maps
    Maße: 243x174x20
    Einband: Taschenbuch
    Reihe: Uni-Taschenbücher L
    ISBN: 9783825284497

    Inhaltsverzeichnis

    Inhaltsverzeichnis
    Vorwort7
    1 Das Leben und seine Bestand-
    teile11
    1.1Der Aufbau von DNA und RNA13
    1.2Der genetische Code16
    1.3Die Gene17
    1.4Die Proteine22
    1.5Die Zelle27
    2Grundlagen der Arbeit im Labor29
    2.1Wasser30
    2.2Messung des pH-Wertes31
    2.3Puffer32
    2.4Waagen33
    2.5Mikropipetten34
    2.6Gefäße im Labor36
    2.7Zentrifugen36
    2.8Mischen und Konzentrieren40
    2.9Konzentrieren41
    2.10Pufferwechsel42
    2.11Proben-Lagerung42
    2.12Steriles Arbeiten43
    2.13 Die kleinen Freunde - Pflege und
    Aufzucht von Escherichia coli45
    2.14Der Aufschluss von Geweben50
    3Aufreinigung von Nukleinsäuren53
    3.1 Gegenspieler erfolgreicher DNA- und
    RNA-Isolationen55
    3.2Extraktion von Nukleinsäuren58
    3.3Die weitere Aufreinigung der DNA59
    3.4 Einige ausgewählte
    DNA-Aufreinigungsmethoden66
    3.5Die Isolation von RNA71
    3.6Übersicht über den Einsatz von Kits74
    3.7 Tipps zum Erzielen hoher Ausbeuten
    bei der Isolation von Nukleinsäuren74
    3.8Quantifizierung von Nukleinsäuren75
    4Polymerase Kettenreaktion79
    4.1 Prinzip der Polymerase-Ketten-
    reaktion81
    4.2Die Komponenten der PCR81
    4.3Geräte für die PCR90
    4.4Die Standard-PCR91
    4.5Ausgewählte PCR-Methoden91
    4.6Anwendungen der PCR100
    4.7Optimierung der PCR-Reaktion101
    5Klonieren für Einsteiger103
    5.1Restriktionsenzyme105
    5.2Ligation113
    5.3Dephosphorylierung115
    5.4Die Transformation von E. coli117
    5.5Der Weg zum Monierten Gen119
    5.6 Der ungerichtete Einbau einer
    amplifizierten DNA in einen Vektor119
    5.7 Der gerichtete Einbau von DNA
    in einen Vektor122
    5.8 Ligase-unabhängige
    Klonierungssysteme124
    5.9Wenn es mal nicht klappt125
    6Vektoren127
    6.1Plasmide129
    6.2Klonierungsplasmide132
    6.3Rekombinase-basierte Klonierung136
    6.4Expressionsplasmide139
    6.5Reportergene143
    6.6Phagen146
    6.7Phagemide147
    6.8Shuttle-Vektoren147
    6.9 Künstliche Chromosomen: BACs,
    PACs und YACs148
    6.10 Hefen als Klonierungs- und
    Expressionssystem149
    .11Die Transformation von Pflanzen151
    .12 Die Transformation von Säugetieren
    und tierischen Zellkulturen153
    7 Elektrophorese von
    Nukleinsäuren157
    7.1 Agarose-Gelelektrophorese
    von DNA159
    7.2Die Detektion der DNA im Gel164
    7.3Präparative Agarosegele165
    7.4 Auftrennen von RNA im
    Agarosegel167
    7.5 Fehlersuche bei der Agarose-
    Gelelektrophorese168
    8Proteinaufreinigung171
    8.1Homogenisation175
    8.2Extraktion von Proteinen177
    8.3Dialyse und Konzentrierung181
    8.4Weitere Aufreinigungsschritte183
    8.5Quantifizierung von Proteinen186
    8.6 Aufreinigungsverfahren für
    getaggte Proteine aus E. coli189
    9 Gelelektrophorese von
    Proteinen195
    9.1 Die Polyacrylamid-Gelelektrophorese
    (PAGE)197
    9.2Die denaturierende SDS-PAGE197
    9.3Das Färben der Gele206
    9.4 Weitere Elektrophoreseverfahren
    für Proteine208
    9.5Western Blotting211
    9.6Wenn es mal nicht klappt214
    10 Immunbiochemische
    Methoden217
    10.1Antikörper219
    10.2 Prinzipien immunbiochemischer
    Verfahren223
    10.3 Spezifische und unspezifische
    Bindungen229
    10.4Immunbiochemische Verfahren230
    10.5ELISA230
    10.6Immunfärbung nach Western Blot235
    10.7Immunoaffinitätschromatographie237
    10.8 Weitere Antikörper-basierte
    Methoden239
    10.9Wenn es mal nicht klappt240
    11 Molekularbiologie für
    Fortgeschrittene243
    11.1DNA-Sequenzierung245
    11.2Bioinformatische Sequenzanalyse248
    11.3 Der Einsatz der Bioinformatik für
    die Klonierung von DNA250
    11.4 Vollständige Sequenzen durch
    3' RACE-PCR und 5' RACE-PCR251
    11.5Stammsammlungen253
    11.6 Identifizierung von Nukleinsäuren
    durch Hybridisierung253
    11.7DNA-Chips oder Microarrays255
    11.8Veränderung von Nukleinsäuren256
    11.9Synthetische Gene260
    Anhang
    Al Sicherheit im Labor262
    A2 Die 10 goldenen Laborregeln263
    A3 Das Laborbuch und die finale Arbeit264
    Verzeichnis der Maps, Protokolle, E-Docs,
    Tabellen und Abbildungen266
    Abkürzungsverzeichnis268
    Quellenverzeichnis28
    Register29
    Alle E-Docs finden Sie im Internet unter http://www.utb-mehr-wissen.de

    Register

    Register
    2-D-Gelelektrophorese 209, 236 3'-OH-Ende 14,81,83,87,98,102,
    107,251 3'UTR 22,251 5'-P-Ende 113,116 5'UTR 22 6xHis-Tag 142f., 19 3f.
    A-Tailing 84,121, 125,
    Acrylamid 201, 203, 214,f.
    Adapterprimer 115, 251f.
    Adenin 12ff., 21
    Adenoviren 129,154f.
    Agarosegel 61, 66, 74f., 78, 81, 84, 91,
    93, 102, 123, 125, 157ff., 165, 167f.,
    197, 204, 246, 253f.
    Agarplatten 44ff., 49, 69, 99, 119, 264
    Agglutinationstest 224, 239
    Agrobacterium tumefaciens 94, 129, 15lf., E-Doc 6-3
    Alkalische Phosphatase 56,145, 227f., 231, 237, E-Doc 10-2
    Aminosäuren 12,16ff., 22ff., 44, 49, 75, 78, 89, 133, 141ff., 175, 186, 199f., 206ff., 220, 229, 250
    - kanonische 24
    - proteinogene 23f.
    Aminosäuresequenz 18,89,229,250
    Ammoniumacetat 63,169
    Ammoniumsulfat 173,179ff., 189, 206,221
    Ammoniumsulfatfällung 179,182, 184
    Ampholyte 22,197,208
    Ampicillin 42, 48f., 133f., 138,149
    Annealing 81, 88ff.
    Annealing-Temperatur 84, 88ff., 102
    Antibiotika 44, 47ff., 119,130, 133
    Antigen 218ff., 259ff.
    Antikörper 9, 42f., 142ff., 153ff., 180, 196, 211, 213,217, 59, 165, 171, 182ff., 187ff., 194, 218ff., 254f., 255, 258f., 264
    - Fänger-A. 233f., 241
    - Monoklonale A. 221
    - Polyklonale A. 218, 221, 225ff., 238
    - rekombinante A. 146, 218, 220f., 258, E-Doc 10-1
    Ausgangs-DNA (=Template DNA) 80ff.,
    93ff., lOOff., 113, 119, 121, 147,
    259f.
    Autoklavieren 36, 43ff., 56, 67,72f.,
    262f.
    ß-Galaktosidase 133ff., 143
    BAC 129, 148ff.
    BCA-Test 33,186ff., 206
    Bibliothek (DNA-, Gen-, Phagen) 222, 257f., E-Doc 11-4
    Bioinformatik 244, 248ff.
    Biotin 142,228,254
    BLAST 248f., E-Doc 11-2
    Blau-Weiß-Screening 120,123,133ff., 139, 143, 149
    Blocklösung 213, 225f., 231ff., 237, 242
    Blotto 226, 232, 234ff.
    Blue Native PAGE PAGE
    Blunt Ends stumpfe Enden Bradford-Test 186ff., 197,178f., 206
    Bromphenolblau 162,164, 199f., 205
    BSA —> Rinderserumalbumin Bunsenbrenner 44f., 262
    CAAT-Box 20
    Calf Intestine Alkaline Phosphatase (CIAP) 116
    Capture Phase 173,233
    Carbenicillin 42,48
    ccc-DNA 132
    CDNA 21, 72ff., 96ff., 119f., 128, 146,
    220, 249, 25Iff.
    CDNA-Bank 71, 73, 97, 253, E-DOC 11-4
    Chaotrope Salze 54, 59, 65f., 72, 125, 169, 179f., E-Doc 8-2
    Chloramphenicol 45, 48f., 145, 149, 151
    Chloroform 54, 56, 60f., 66, 69f., 72, 167
    Chromatin Immunopräzipitation (ChIP) 224
    Chromatogramm - Protein 183, 265, E-Doc 8-3
    - DNA 245ff.
    Chromatographie 74,172f., 182ff., 193ff., 217ff., 226, E-Doc 8-4
    - Affinitäts-C. 184ff., 219, 221, 237f.
    - Gelfiltrations-C. (SEC) 184, 215
    - Hydrophobe Interaktionschromato- graphie (HIC) 184
    - Immunoaffinitäts.-C. 237f.
    - lonenaustausch-C. (IEX) 184,221
    - Metallchelatchromatographie
    (IMAC) 194f.
    Cl-Methode (s.a. PCI) 60f., 69
    codierender Strang 18
    codogener Strang 18
    Codon 16f., 24, 89, 141, 144, 250f., 257
    Codon-Optimierung 144, 257
    Codon-Usage (Tabellen) 251,257
    Cofaktor 26, 57,176
    Coomassie Brilliant Blue 186, 206ff.
    Coomassiefärbung 191,196f., 205, 206ff., 236
    Cosmid 129, 148f CTAB 58f., 64f., 69, 71, 125, 169, E-Doc 3-1
    Cy3, Cy5 254ff.
    Cytosin 12ff.
    Dam-Methylase 111, 112f., 126, 260
    dcm-Methylase 112f., 126
    DEPC (Diethylpyrocarbonat) 56, 72f., 98
    Desoxynukleotid (dNTP) 13, 63, 80f., 86f., 98, 102, 123, 147, 244f., 254
    Desoxyribonukleinsäure (DNA) 12
    Dialyse 42, 66,181f., 215
    Didesoxynukleotid (ddNTP) 244f.
    Diethylpyrocarbonat -> DEPC
    Digoxygenin 254
    Dimethylsulfoxid 49, 86,102, 167, 175
    Display Verfahren Phage Display Disulfid-lsomerase 141,193
    Dithiothreitol (DTT) 58f., 69f., 98, 111, 115, 176, 178, 189, 199, 214
    DMSO Dimethylsulfoxid
    DNA-Chips 99, 255f.
    DNA-Microarray 71f., 99, 244, 253ff.
    DNA-Polymerase —> Polymerase DNA-Sequenzierung -> Sequenzierung DNase 55f., 58f., 105, 175
    DNase I 51,73
    dNTPs —>• Desoxynukleotid Dot Blot 211
    Dpnl 107f., 113, 259
    Dpnl-Mutagenese —> Mutagenese
    Drigalskispatel 44f., 119, 212f.,
    dsRED 144f.
    Durchflussphotometer 183,186
    Durchflusszytometrie 240
    EDTA 55, 57f., 66, 68ff., 78, 84, 86, 101, 16 If., 182, 189
    EDTA-Blut 70
    Einschlusskörperchen 142,192
    Elektrokompetente Zellen 117f., 128
    Elektroporation 117ff., 150f.
    ELISA 9, 34, 143, 218, 224, 226, 230ff.,
    240f.
    - kompetitiver ELISA 233ff.
    - Sandwich ELISA 233f., 241
    Elongation 80, 83, 249
    Episom 49,130,133,150
    Epitop 218, 22Of., 223, 229f., 233f.
    Escherichia coli 30, 45ff., 49, 51f., 59, 66, 68, 83, 101, 105, 112, 117, 119, 126, 129f., 13 6f., 139ff., 143, 147ff., 154ff., 176, 190ff., 222, 246, 257, 259f., E-Doc 2-7
    EST-Bank 97f., E-Doc 11-4
    Ethanolfällung 56, 61ff., 68f., 73,102, 168, 177, 247
    Ethidiumbromid 132,162,164ff., 262
    Exon 20f., 97
    Exonuklease 55f., 84f., 100, 124, 251
    ExoSAP-IT 116, 247, E-Doc 5-2
    Expression - heterologe E. 9,21,45,97,139, 153, 155, 190ff., 233, 257
    - homologe E. 139
    Expressionsvektor 110,128f., 130, 139f., 142, 190f., 249
    Anhang Extension Elongation F Plasmid / F* Plasmid -> Plasmid FACS 240
    FASTA 249, E-Doc 11-2
    Flag-Tag 142,237
    Fluorescein 228,239,254
    Fluoreszenzfarbstoff 89,100,145, 196, 224, 227f., 240, 244f., 255ff., E-Doc 9-2
    Fluoreszenzmikroskopie 224, 228, 239
    Fluoreszenzprotein 144f.
    Fluorometer 78, E-Doc 3-5
    Flüssiger Stickstoff 49ff., 58, 64, 69,174
    Freeze-'n'-Squeeze 166f.
    Freeze-Thaw 52
    Fusionsprotein 143,155,190, 258f.
    Gateway-System 130,137f.
    GC-Gehalt 15, 77, 89,102, 254
    genomische DNA 52,56,71,80,89,
    102, 119, 148, 253f.
    Gewebekultur 45, 50,174, 176
    GFP (Green Fluorescent Protein) 143ff., 151f., 155
    Glasmilch 65, 74
    Glucose 47,67,140,189
    Glycerin 43, 46, 49,102,112,117,126, 162,199, 204
    Glykosylierung 17, 28
    Gradientencycler 90, 92
    Guanidinsalze 59, 65f., 72, 189
    Guanidinthiocyanat (GTC) 56,59,72
    Guanin 12ff., 20
    m7G-Cap 2Off., 252f.
    Hairpin 88f., 123
    Harnstoff (Urea) 101,190,192
    Heavy chain -> Schwere Kette Hefe -> Saccharomyces Heidelberger Kurve 223
    High-Copy-Plasmid Plasmid
    Hitzeschock-Transformation 117,
    119f., 126,173f.
    Homogenisation 37, 50, 52, 55f., 58f., 71f., 74, 174ff., 185
    Homogenisationspuffer 50f., 55, 58, 69f., 176, 178,199
    Hybridisierung 81,158, 167f., 244,
    253ff.
    Hybridoma 154f., 218, 221
    Hypochromer Effekt 15,77
    Immunfärbung 9, 40f., 143,189, 21 Off., 224ff„ 235ff., 239ff„ 255, 265
    Immunfluoreszenz 224,226,239
    Immunfluoreszenz-Mikroskopie 228
    Immunisierung 221
    Immunglobulin Antikörper Immunogen 219,221
    Immunpräzipitation 219,223f., 227
    Inclusion Bodies -> Einschlußkörper- chen Inkompatibilitätsgruppe 130
    Inkubationsschüttler 40f., 46,191
    Intron 20f., 97, 144,152,156
    IPTG 134ff., 140, 142, 190f.
    Isoelektrische Fokussierung 196f.,
    208f.
    Isoelektrischer Punkt 22, 24,181, 197,
    208ff.
    Isopropanol 64,68,70,73,169
    Isoschizomer 109f., 112
    Kanamycin 48f., 138,151,155
    Kapillarelektrophorese 244f.
    Kit 59, 66, 71, 73f., 84, 136, 146, 164, 166f., 189, 194, 233, 246f., 253, 259, E-DOC 3-4
    Klonierungsvektor 129ff., 148ff.
    Knockout-Maus 153, E-Doc 6-4
    kohäsiven Enden 108, 113,119,129
    kompetente Zellen (Ca2+) 117, 126, E-Doc 5-3
    Kugelmühle 50f., 174
    lacZ 47, 133ff., 139f., 145, 148f., 190f.
    lacZ-Promoter-> Promoter LB-Medium 46f., 136,191
    Leichte Kette (LC) 219f.
    Leupeptin 175,192
    Ligase-unabhängige Klonierungs- systeme (HC) 120,124, Ligase 75, 95,105,113ff„ 120f., 126, 131, 136, 177
    Ligation 81,113ff., 117ff., 123, 126
    Low-Copy-Plasmid -> Plasmid Low-Melting Agarose 160
    Low ry-Test 186,188f.
    Luminol 227
    Lysozym 51, 59,174,192f.
    M13-Phage —> Phagen Magnetic Beads 74,184ff., 193, 226
    Magnetrührer 41,179,181f.
    Massenspektrometrie 196,210,250, E-Doc 8-6
    Mastermix -> PCR-Mastermix
    Matrizen-DNA Ausgangs-DNA
    Maus -> Mus musculus Meerrettich Peroxidase (HRP) 227f., 231, 234, 242, E-Doc 10-2
    Methylase 105ff., 112f., 126
    Microarray DNA-Microarray Mikropipette 34f., 57, 232, 264
    Mikrotiterplatte 37,188, 230ff., 241, 246
    Minizentrifuge —> Zentrifuge Mörser 50,58,69,174,177
    Multiple Cloning Site (MCS) 133ff., 139f., 156
    Mus musculus (Maus) 153,155,221, 253
    Mutagenese 90,100,113, 256, 258f.
    - Dpnl-basierte M. 259f.
    - Error Prone M. 259
    - gerichtete M. (Site-directed M.) 257
    - Saturation M. 257
    - Scanning M. 257
    NanoDrop 76
    native PAGE PAGE
    Natriumacetat 32, 63ff., 67f., 86, lOlf., 123, 169, 189
    Ncol 109f., 112
    Neomycin 48f., 155
    Neoschizomer 110
    Nitrozellulose 211f., 226, 239
    Northern Blot 71ff., 158,167f., 211, 244, 253, 255, E-Doc 7-2
    Nuklease 55ff., 70, 95,105,107, 126, 147,168
    Nukleotid 12ff., 63, 80, 84ff., 90, 108f., 114, 121, 141, 244, 250, 259
    Nylonmembran 255
    Oberflächensterilisation 45
    occ-DNA 132
    Oligonukleotid (s.a. Primer) 63, 65, 77,
    89, 100, 115, 253f.
    One Hybrid System 257
    Operon 18f., 148
    ori —> Replikationsursprung
    PAC 129,148ff.,
    PAGE (Polyacrylamidgelelektrophorese)
    197ff.
    - Blue Native PAGE 208
    - native PAGE 196f., 208, - SDS-PAGE 172f., 178, 181f., 189, 191, 196ff., 205, 208ff., 213f., 218, 236, 242, E-DOC 9-1
    Panning 259
    Paratop 221, 223, 229f.
    PBS (Phosphate-Buffered Saline) 42,
    222,226, 231, 234, 236ff.
    PCI 56, 6Of., 69,71f., 167
    PCR (Polymerase-Kettenreaktion) 8,
    31, 36ff., 68, 70, 77, 80, 81ff., 104f.,
    113, 115, 119ff., 136ff., 159, 163,
    166, 168, 224, 231, 245ff., 250ff.,
    259f.
    - Error Prone PCR 259
    - Gradienten-PCR 90,92
    - inkrementelle PCR 92,102
    - Kolonie-PCR 82, 84, 99,119f., 123, E-DOC 4-2
    - Multiplex-PCR 93f.
    - Nested PCR 92f., 102, 251
    - Quantitative PCR lOOf., E-Doc 4-3
    - RACE-PCR 25Iff.
    - RT-PCR 71, 73, 96ff., 100, 168,
    25 If.
    - Sticky-End PCR 95, E-Doc 4-1
    PCR-Puffer 86,91,99
    PCR-Mastermix 91, 99f.
    pCRScript 12lf., 134, 251
    PEG -> Polyethylenglykol PEG-Fällung 64, 68
    Peiß-Sequenz 141,193
    Periplasma 48,141,175,190,192f.
    pH-Meter 31f.
    pH-Indikator 61, 73, 162,199f.
    pH-Wert 22, 24, 30, 31ff., 46f., 63, 66f., 78, 176, 178, 181, 201, 208f., 222, 238, 242, E-Doc 2-1
    Phagemid 59,129,131, 136,147f., 222, 259
    Phagen 36, 45, 59, 64, 105f., 129ff.,
    146ff., 222, 258f.
    - Phage fl 147,155
    - Phage A 114,129,136f., 140, 146ff., - Phage M13 129, 146f., 258, E-Doc 11-5
    - Phage PI 129,148f.
    - Phage T7 191
    Phage Display 129,141, 146f. 218, 245, 258f., E-Doc 11.5
    Phenol 54, 56, 58, 60f., 70, 72,101, 167, 176
    Phenolextraktion PCI
    Phenoloxidase 58,174,176
    Phosphatpuffer (s.a. PBS) 32, 56, 63, 177f., 222
    Photometer 46, 67, 75ff., 100,186, 188, 231
    Pipette Mikropipette Pipettenspitze 34ff., 44f., 69f., 99,102
    Plasmid 9, 26, 42, 57, 59, 66ff., 74, 81,
    89, lOlf., 104f., 118ff., 124, 129ff.,
    138ff., 147ff., 163, 246, 253, 259f.
    - 2p Plasmid 150
    - binäres Plasmid 151f.
    - Broad Range Plasmid 147,152
    - Centromer-haltige Plasmide 150
    - Episomale P. 150
    - Expressions-P. 110, 130,139f.,
    190f.
    - F Plasmid 129ff., 148f.
    - F' Plasmid 133ff., 140, 148
    - Helferplasmid 151
    - High Copy P. 130, 133
    - Integrative P. 150
    - Klonierungs-P 129ff. 132ff., 139, 147,151
    - Low Copy P. 130, 139, 246
    - Ti-Plasmid 151f.
    Plasmidisolationskit 69, 247
    Plasmidpräparation 59, 66ff., 102,
    112, 163, 191, 246f.
    PMSF 175, 192, 197
    Polishing Phase 173,182
    PolyA+-RNA 22, 61, 71, 97f., 119, 185, 251, E-Doc 3-3
    Polyacrylamidgelelektrophorese
    PAGE
    Polyadenylierung 21f., 74
    Polyadenylierungssignal 21, 152, 155f., 251
    polycistronisch 19
    Polyethylenglykol (PEG) 61, 64, 68,
    181f.
    Polymerase 18
    - DNA-P. 18, 56, 73, 80ff., 83f., 95ff., 121, 123f., 147
    - Proofreading DNA-P. 84f., 120f., 123
    - reverse DNA-P. reverse Transkrip- tase - RNA-abhängige DNA-P. 97
    - RNA-P. 18ff., 140,191
    - Taq-DNA-P. 56, 83f., 121f.
    - T7-RNA-P. 140, 19 Of.
    - Tth P. 97
    Polymerase-Kettenreaktion -> PCR
    Polynukleotid Adenylyltransferase 56, E-DOC 5-5
    Polyphenol 57f.,70, 102, 176
    Prä hybridisierung 255
    Primer 80f., 83f„ 87ff., 119ff., 132, 134, 138, 140, 149, 166, 168, 245ff., 249ff., 259
    - Adapterprimer 251f.
    - degenerierte P. 87, 89f., 92,119,
    2 5 Of.
    - Hexamer-P. 97,99
    - modifizierte P. 89
    - nested Primer 93
    - Oligo-dT-Primer (= oligio-dT18.22VN- Primer) 98, 251f., 252
    - Primerdimer 89,96,102,123
    - Sequenzierprimer 244ff.
    - Smart™-Primer 252f.
    Probenpuffer
    - für DNA 162f.
    - für SDS-PAGE 198f., 214f Promoter 18ff., 47,132ff., 139, 140ff., 143, 152, 154f., 190f., 257
    - 35S-Promoter 152
    - Early-SV40-Promoter 155f.
    - lacUV5-Promoter 140
    - lacZ-Promoter 47,133ff., 139f.,
    19 Of.
    - nos-Promoter 152
    - PL-Promoter 140
    - T7-Promoter 139f., 190,191
    Promoteranalyse 143,145, 257
    Proofreading Polymerase Polymerase Proteaseinhibitor 51,175f., 199
    Protease 58,139, 173ff., 192, 194, 214, 220, E-Doc 8-1
    Proteinase K 58f., 70
    Protein Engineering 258
    Proteinsynthese 19, 27f., 49
    Pufferkapazität 31f., 168
    Purin 12ff.
    PVDF-Membran 211ff., 226, 237
    Pyrimidin 12ff.
    Ouartärstruktur 25f.
    RACE 7If., 97, 249f„ 251ff.
    rbs -> ribosomale Bindestelle Reaktionsgefäß 31, 35ff., 49, 63, 68f., 74, 9Of., 96, 99f., 124, 166f., 185, 188, 264
    Red Safe 165
    Reinstwasser 31, 66,118
    Reinstwasseranlage 30f., 66
    Rekombinase 120, 124,128,136ff., - cre-Rekombinase 149
    Rekombinase-basierte Klonierung
    136ff.
    Rekombinase-Site 137f., 149, 156
    Rekombination 130,136ff., 150
    - homologe R. 150f., 154, 257
    - illegitime R. 151
    Renaturierung 15, 59, 67, 88
    Replikation 13
    Replikationsursprung 128, 130, 132ff., 139, 146f., 149ff., 156
    - oriColEl 130
    - pSa-ori 151f.
    - pUC-ori 155
    - SV40-ori 150, 155
    Reportergen 143f., 152,155, 257
    Restriktionsenzym 57, 95f., 105ff., 119ff., 126, 132f., 136f., 163, 166, 177, 254, 259
    Retroviren 12, 17, 96, 129, 154
    Reverse Genetik 153,245,257
    Reverse Transkriptase 21, 96ff., 119, 253,255
    - AMV R.T. 96
    - M-MLV R.T. 96, 98
    Reverse Transkription 12, 72, 96f., 119f., 249, 253
    Ribonukleinsäuren —>• RNA
    Ribosom 19
    ribosomale Bindestelle (rbs) 19, 139, 141
    Ribozym 16, 18f., 22, 26f., 28, 40, 49
    Rinderserumalbumin (BSA) lllf., 187f., 226
    - deacetyliertes BSA 102
    R-M-System 106, 112
    RNA 12ff., 52, 55ff., 63, 65f., 70ff., 85, 87, 96ff., 110, 115, 119f., 139, 146, 158, 167f., 177, 185, 251, 253ff., 257
    - hnRNA 2Of.
    - mRNA (messenger RNA) 12,16, 18ff., 27, 49, 54, 73, 96f., 99f., 141, 156, 249, 25Iff., 254, 256, 258
    - rRNA (ribosomale RNA) 16,18,22,
    40, 73, 167f.
    - tRNA (Transfer-RNA) 16, 18f., 22, 49, 73, 257
    RNA-Ligase 252
    RNA-Polymerase —> Polymerase RNA-Sonde 139
    Anhang RNase 25, 56, 67ff., 98
    - RNase A 68
    - RNase H 69,96,98
    - RNase Block 98
    RNase-freie DNase 73
    Rolling Circle 147f.
    Rotationsmischer 40f.
    Rotor 37ff., 41
    Saccharomyces cerevisiae (Hefe) 59, 129, 148, 150f., 253, 257, E-Doc 6-2
    Sammelgel 202f., 215
    Säuger-Zellkulturen 150,153ff., 257
    scFv —> Antikörper, rekombinant Scherkraft 40f., 41, 52, 55ff„ 71,126, 131
    Schmelztemperatur 15, 77, 88f., 255
    Schüttler 40f., 46, 70,119, 191
    Schwere Kette (HC) 219f., 258
    Screening (s.a. Biau-Weiß-Screening) 99, 120, 134, 258
    SDS (Natriumdodecylsulfat) 30,42,
    58f., 63, 67f., 70ff., 101, 178, 189,
    197, 199f., 203ff., 208, 212, 214f.
    SDS-Gelelektrophorese PAGE
    SDS-PAGE PAGE
    SDS-Laufpuffer 204,215
    SDS-Probenpuffer 198
    Sekundärstruktur 24
    Selbstligation 115f.
    Selektion 47,128,133,152,258
    Selektionsverfahren 258
    Self-Priming 89
    Semi-Dry Blot 211ff., 216
    Sequenzierung - DNA-S. 66, 88, 132, 134, 146, 244ff., E-DOC 11-1
    - Protein-S. 252, E-Doc 8-7
    Shine-Dalgarno-Sequenz 18, 139,141
    Shrimp Alkaline Phosphatase 141
    Shuttle-Vektor 129,147f., 150f., 154ff.
    Silberfärbung 206ff., 214
    Silika-Partikel 65f., 91,121,123,166f.
    Southern Blot 113,158,211,244, 253ff., E-Doc 7-1
    Speed-Vac 41f., 57, 62, 64, 68,101,198
    Speedcycler 90
    SpektralphotometerPhotometer Spheroblasten 150
    Spleißen 20f.
    - alternatives S. 20f Srfl 110,121f., 134
    Srfl-basierte Verfahren 120
    ssDNA 14f., 75, 77, 114, 129, 146f., 245, 254
    Stain G 165f.
    Star-Aktivität Ulf., 126
    Startcodon 17,19, 21f., 110, 141
    Sterilbank 31, 44f., 135f.
    Sticky ends —> kohäsive Enden Stoppcodon 17,19, 21f., 110,141
    Streptavidin 142, 228, 254
    Stuffer-DNA 146,149
    stumpfe Enden (blunt ends) 105,108, 113ff., 119, 121f., 124, 136,251
    supercoiled DNA 74,125,131f., 163
    SYBR Green 100
    T7-Promoter Promoter
    T7-RNA-Polymerase —> Polymerase
    Tag-Sequenzen 98,139f., 141,142ff.
    145, 184, 190, 192ff.
    T-DNA 15 Iff.
    T-Antigen 155f.
    T4-Ligase Ligase TA-Klonierung 80, 84,120f., 125, 251
    TAE-Puffer 42, 56ff., 160ff., 164, 167ff.
    Taq-Polymerase Polymerase TATA-Box 20
    TBE-Puffer 56ff., 161
    TBS-Puffer 222,238
    TCA-Fällung 181,198,200,213
    TE-Puffer 57, 62, 65f., 68ff., 75, 86, 91, 101, 167
    Template-DNA -> Ausgangs-DNA
    Tertiärstruktur 25,163,179
    Tetracyclin 48f.
    Thermocycler 80f., 83, 90ff., 100,115
    Thymin 12ff.
    tierische Zellkultur Säuger-Zellkultur Tissue Print 239
    TOPO-Cloning 125
    Topoisomerase I 124f., 131f., 137
    Transduktion 129, 156
    Transfektion 156
    Transkriptionsfaktoren 20f.
    Translation 12,16ff., 19f., 22, 24, 26f., 49, 56, 85, 141, 257
    Trenngel 202f., 215
    Transformation 8,94,99,105,117, 117ff., 123f., 126, 133, 144, 147, 259
    - von Hefen 151
    - von Pflanzen 129,151f.
    - von Säugerzellen 153ff.
    Transkription 8,12,16ff., 26, 80, 255
    Tris-Puffer (s.a. TE-, TBE- und TBS- Puffer) 33,73
    Trockeneis 51, 64
    TY-Medium 46f., 117,191
    Ultrafiltration 41,182
    Ultraschall 50, 51f., 174,192f., 224
    Ultrazentrifuge Zentrifuge
    Uracil 12f., 15ff.
    UV-Crosslinking 255
    UV-Tisch 164ff., 255
    UV-Photometer Photometer Vektoren (s.a. Plasmid) 9, 80, 95, 97, 104f., 114ff., 119ff., 128ff., 140f., 144f., 147f., 150f., 154ff. , 166, 245
    - binärer V. 150ff.
    - BAC-V. 148
    - Entry-V. 13 7f.
    - Expressions-V. 110,128f., 130, 139f., 142, 190f., 249
    - Klonierungs.-V. 129ff., 148ff.
    - PAC-V. 148f.
    - Replacement-V. 146
    - Shuttle-V. 129, 147f., 154ff., - Suicide-V. 122, 136
    - T-Vektor 121
    Vektorkarte 141
    Vortexer 40f., 49, 57, 62, 71
    Waage 33
    Wasser 12, 30ff., 41f., 45, 60, 71, 73, 117, 161, 168, 174,180f., 193, 236
    Wasserbad 51, 69, 112,117,166,198f.
    Wasserkontrolle 83, 93f., 101f.
    Wasserstoffbrücke 14f., 25, 77, 87, 229
    Wasserstoffperoxid 227
    Western Blot 44, 203, 208, 211ff., 218, 224, 226, 228, 235f., 240, 242, 255
    Wet Blot 211,214,216
    Wöbbel 16, 90, 250f.
    Xbal 109f., 126
    X-Gal 133ff., 145
    Xylencyanol 162
    YAC 56,129,148,150
    Zentrifuge 36ff., 42, 51, 63f., 167 178ff., 192, 198, 236, 262, 265
    - Kühlzentrifuge 37ff.
    - Minizentrifuge 38f., 91
    - Ultrazentrifuge 39
    Zwitterion 22, 24, 178
    Zymolase 59