Methods and techniques in virology:
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
New York u.a.
Dekker
1993
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XVII, 309 S. Ill., graph. Darst. |
| ISBN: | 0824791010 |
Internformat
MARC
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| 245 | 1 | 0 | |a Methods and techniques in virology |c Pierre Payment and Michel Trudel ... |
| 264 | 1 | |a New York u.a. |b Dekker |c 1993 | |
| 300 | |a XVII, 309 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 650 | 7 | |a Virology |2 cabt | |
| 650 | 7 | |a Techniques |2 cabt | |
| 650 | 4 | |a Virology |x Technique | |
| 650 | 4 | |a Virology |x methods | |
| 650 | 0 | 7 | |a Methode |0 (DE-588)4038971-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Virologie |0 (DE-588)4063597-1 |2 gnd |9 rswk-swf |
| 689 | 0 | 0 | |a Virologie |0 (DE-588)4063597-1 |D s |
| 689 | 0 | 1 | |a Methode |0 (DE-588)4038971-6 |D s |
| 689 | 0 | |5 DE-604 | |
| 700 | 1 | |a Payment, Pierre |e Sonstige |4 oth | |
| 700 | 1 | |a Trudel, Michel |e Sonstige |4 oth | |
| 856 | 4 | 2 | |m HEBIS Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=005513855&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-005513855 | ||
Datensatz im Suchindex
| _version_ | 1804122741216903168 |
|---|---|
| adam_text | Methods and
Techniques in
Virology
Pierre Payment and Michel Trudel
Institut Armand-Frappier
University du Quebec
Ville de Laval Qu6bec, Canada
with contributions by
Robert Alain, M Sc
Max Arella, Ph D
Serge Belloncik^ Ph D
Laurent Berthiaume, Ph D
Andr6 Fafard, M Sc
Francois Fossiez, Ph D
Claude Hamelin, Ph D
Michel Houde, Ph D
Patricia Jouvenne Ph D
Jacqueline Lecomte, Ph D
Gilles Lussier, DMV, Ph D
Claire Simard, Ph D
Pierre Talbot, Ph D ,
J Arnold Verbeek, Ph D
Marcel Dekker, Inc New Ybrk • Basel • Hong Kong
Contents
1 LABORATORY SAFETY 1
1 INTRODUCTION 1
2 TYPES OF CONTAINMENT 3
2 1 Appropriate microbiological practices 3
2 2 Biosafety equipment 4
2 3 Installations 4
3 BIBLIOGRAPHY 4
2 MONOLAYER CELL CULTURE 5
1 CELL CULTURE MEDIA 5
1 1 Use of Antibiotics 6
1 2 Preparation, sterilization and conservation 7
1 3 Cell-dispersing solutions 8
131 Use of dispersing solutions 9
1 4 Dispersion of cell monol ayers 9
1 5 Preparation of primary cell cultures 11
151 Kidney primary culture 11
152 Avian fibroblast cultures 12
2 MAINTENANCE OF CELL CULTURES 12
2 1 Primary cell cultures 12
2 2 Continuous cell lines 12
2 3 Maintenance of diploid cell lines 13
231 Culture from frozen cells 13
232 Subculture of diploid cells 14
2 4 Enumeration of cells 14
241 Crystal violet staining 14
242 Trypan blue staining 15
3 CELL CLONING 15
4 COLONY FORMATION TEST 16
5 CRY OPRESERV ATION 17
5 1 Freezing of cells 17
5 2 Recovery, of frozen cells 17
6 CHROMOSOME PREPARATIONS 17
7 REFERENCE 18
3 ISOLATION AND IDENTIFICATION OF VIRUSES 19
1 PREPARATION OF SPECIMENS 19
1 1 Stool samples 19
1 2 Swabs and biological fluids 20
1 3 Tissues 20
1 4 Suckling mice organs 20
2 INOCULATION TO EMBRYONATED EGGS 20
2 1 Amniotic route 20
2 2 Allantoic route 22
2 3 Yolk sac route 22
2 4 Intravenous route 22
2 5 Chorioallantoic membrane route 22
v
vi Contents
3 ISOLATION OF VIRUSES IN CELL CULTURES 23
3 1 Choice of cell cultures 23
3 2 Virus isolation 23
321 Macrotechnique 23
322 Microtechnique 24
4 IDENTIFICATION OF VIRUSES 25
4 1 Hemagglutination 25
411 Buffers 25
412 Preparation of red blood cells 27
413 Hemagglutination characteristics of some viruses 27
414 Titration of a hemagglutinating virus 29
4 2 Hemadsorption 29
421 Virus detection 29
422 Hemadsorption-inhibition test 30
4 3 Cytopathic effect 30
431 Titration of viruses in cell culture by cytopathic effect 32
432 Determination of virus titer by infectivity 33
4321 Reed-Muench method 33
4322 Karber method 33
4 4 Plaque assay 33
441 Enterovirus 33
442 Other viruses 34
4 5 Titration by interference 35
4 6 Neutralization of infectivity 36
4 7 Sensitivity to ether 36
4 8 Sensitivity to 5-IUDR 37
4 9 Electron microscopy 37
4 10 Temperature sensitivity test 37
4 11 Inoculation of newborn mice 38
5 REFERENCE 38
4 ANIMAL MODELS 41
1 INTRODUCTION 41
2 MAJOR SPECIES 41
2 1 Mouse 41
2 2 Rat 42
2 3 Guinea pig 43
2 4 Hamster 43
3 FACTORS INFLUENCING THE CHOICE AND USE OF
ANIMALS 43
3 1 Factors related to the host 43
311 Genotype 43
312 Age 43
313 Sex 44
314 Microbiological status 44
315 Physical environment 45
3 2 Factors related to the agent 45
321 Virulence 45
322 Dose 46
323 Inoculation route 46
Contents vii
3231 Intracerebral route 46
3232 Peripheral routes 46
32321 Respiratory route 46
32322 Oral route 47
32323 Intravenous route 47
32324 Intraperitoneal route 47
32325 Subcutaneous and intradermal
routes 47
32326 Intrauterine route 47
4 VISIBLE SIGNS 47
4 1 Clinical Symptoms 47
4 2 Lesions 48
4 3 Serology 48
4 4 Viral isolation 49
5 BIBLIOGRAPHY 49
5 INOCULATION OF LABORATORY ANIMALS 51
1 INTRODUCTION 51
2 SUPPLIES 51
2 1 Syringes 51
2 2 Needles 51
3 METHODS 52
3 1 Intraperitoneal inoculation 52
3 2 Intravenous inoculation 52
3 3 Intracerebral inoculation 53
6 CONCENTRATION OF VIRUSES 55
1 INTRODUCTION 55
2 METHODS 56
2 1 Hydroextraction 56
2 2 Precipitation 56
221 Precipitation with polyethylene glycol 56
222 Precipitation with ammonium sulfate 57
2 3 Molecular filtration 57
231 Stirred cells 57
232 Thin-channel and tangential flow systems 58
233 Hollow-fiber systems 58
3 APPLICATIONS 58
4 BIBLIOGRAPHY 60
7 PROTEIN ASSAYS 61
1 INTRODUCTION 61
2 SPECTROPHOTOMETRY METHOD 61
3 COLORIMETRIC METHOD 62
3 1 Lowry assay 62
3 2 Dye binding assay 64
321 Standard test (25-125 p g and 0 25-1 25 mg/ml) 64
322 Modified standard test (25-125 ng/ml) 65
323 Microtest (1-20 ng and 1 25-25 ng/ml) 65
324 Microtest in microtiter plates (0 5-5 |Xg and 5-50 jig/ml) 65
viii Contents
3 BIBLIOGRAPHY 66
8 ULTRACENTRIFUGATION 67
1 INTRODUCTION 67
2 DIFFERENTIAL CENTRIFUGATION 67
3 DENSITY GRADIENT CENTRIFUGATION 68
3 1 Centrifugation media 71
3 2 Types of gradients 75
3 3 Types of rotors 75
3 4 Tubes 76
341 Ultra-Clear™ tubes 76
342 Polyallomer tubes 76
343 Polycarbonate tubes 78
3 5 Density gradient fractionation 78
4 PARAMETERS OF ULTRACENTRIFUGATION 80
4 1 Sedimentation coefficient 80
4 2 Buoyant density 80
43K and K factors 80
4 4 Samples 80
5 BIBLIOGRAPHY 81
9 CHROMATOGRAPHY 83
1 INTRODUCTION 83
2 GEL FILTRATION 83
3 ION-EXCHANGE CHROMATOGRAPHY 85
4 AFFINITY CHROMATOGRAPHY 86
4 1 Strategy 86
4 2 Preparation of inhibitor 86
4 3 Preparation of adsorbent 86
4 4 Solubilization of viral proteins 88
4 5 Chromatographic separation 88
4 6 Analysis of fractions 89
4 7 Isolation of hemagglutinins 89
4 8 Results 89
481 Neuraminidase isolation 89
482 Hemagglutinin isolation 89
5 CONCLUSION 91
6 BIBLIOGRAPHY 91
10 ELECTROPHORESIS 93
1 INTRODUCTION 93
2 METHOD 95
2 1 Preparation of gels 95
2 2 Preparation of specimens 96
2 3 Electrophoresis 96
2 4 Coomassie Blue staining 96
2 5 Silver staining 97
3 DISCUSSION 97
4 BIBLIOGRAPHY 98
Contents ix
11 ELECTRON MICROSCOPY 99
1 INTRODUCTION 99
2 SPECIMEN FIXATION 99
2 1 Fixation 100
211 Osmium tetroxide 100
212 Potassium permanganate 101
213 Aldehyde-based fixatives 101
2 2 Dehydration 101
2 3 Embedding media 101
231 Epoxy resins 102
232 Polyester resins: Vestopal W 102
233 Water-soluble embedding media 102
234 Methacrylate 102
2 4 Embedding with Vestopal W 103
2 5 Ultrathin sectioning 103
251 Cryotomy 103
2 6 Staining of sections 104
261 Uranyl acetate 104
262 Lead citrate 104
3 NEGATIVE STAINING 104
3 1 Direct staining on grid 105
3 2 Freezing and thawing method (cryolysis) 106
3 3 Osmolysis - 106
3 4 Concentration on agar 106
3 5 Purified virus suspension 106
3 6 Micro-ultracentrifugation 107
4 BIBLIOGRAPHY 109
12 SEROLOGY 111
1 SUPPLIES Ill
1 1 Multiwell plates Ill
1 2 Calibrated pipettes Ill
1 3 Microdiluters 112
1 4 Verification of micropipettes 112
1 5 Centrifuge carriers for plates 112
2 HEMADSORPTION-INHIBrriON TEST 112
3 HEMAGGLUTINATION 113
3 1 Preparation of hemagglutinating antigens 113
311 Embryonated egg fluids 113
312 Brains of newborn mice 113
313 Cell culture fluids 113
3131 Freeze-thawing 113
3132 Concentration of culture supernatants 114
31321 Ammonium sulfate precipitation 114
31322 Polyethylene glycol precipitation 114
3 2 Treatments to unmask hemagglutinins 114
321 Alkaline buffer extraction 114
322 Tween 80-ether treatment 114
3 3 Titration of hemagglutinins 115
x Contents
4 HEMAGGLUTINATION INHIBITION (HI) TEST 115
4 1 Treatment of sera 115
411 Acid kaolin 115
412 Heparin-manganese chloride 115
413 Trypsin 115
414 Receptor-Destroying Enzyme 116
415 Acetone treatment 116
416 Adsorption of nonspecific agglutinins 116
4 2 Hemagglutination inhibition test 117
4 3 Applications 118
431 Rubella virus 118
4311 Treatment of sera 118
4312 Hemagglutination (titration of the antigen) 118
4313 Hemagglutination inhibition test 119
432 Other viruses 119
5 NEUTRALIZATION TEST 121
5 1 Diagnosis of an infection 121
511 Screening 121
512 Neutralization test 123
6 IMMUNOASSAYS AND DETECTION OF SPECIFIC CLASSES OF
IMMUNOGLOBULINS 123
7 BIBLIOGRAPHY 123
13 IMMUNOFLUORESCENCE 125
1 INTRODUCTION 125
2 PREPARATION OF MATERIAL 126
2 1 Viruses with rapid CPE 126
2 2 Viruses without CPE 127
2 3 Fixation of cells or tissues 127
2 4 Preparation of tissues 127
241 Cryomicrotomy 127
242 Paraffin embedding 128
3 METHODS 128
3 1 Direct iifimunofluorescence 128
3 2 Indirect immunofluorescence 130
3 3 Elimination of non-specific fluorescence 130
4 MICROSCOPY 130
5 BIBLIOGRAPHY 131
14 IMMUNOPEROXIDASE 133
1 INTRODUCTION 133
2 LIGHT MICROSCOPY 134
2 1 Fixation of sample 134
211 Smears and cell monolayers 134
212 Cell cultures in microplates 134
213 Tissue sections 134
2 2 Direct method 134
2 3 Indirect method 135
3 PAP AND ABC METHODS 136
4 BIBLIOGRAPHY 136
Contents xi
15 ELISA 139
1 INTRODUCTION 139
2 ANTIBODY DETECTION 140
2 1 Preparation of viral antigens 140
2 2 Direct method 140
2 3 Indirect method 142
2 4 Expression of ELISA titers 142
3 BIBLIOGRAPHY 142
16 MONOCLONAL ANTIBODIES: IDENTIFICATION OF
ANTIGENIC SITES 143
1 INTRODUCTION 143
2 COMPETITION ASSAY 144
2 1 Coupling immunoglobulins to peroxidase 144
2 2 Biotinylation 145
2 3 Competition assay 145
3 SELECTION OF ANTIGENIC VARIANTS 146
4 BIBLIOGRAPHY 148
17 HISTOLOGY AND HISTOCHEMISTRY 151
1 INTRODUCTION 151
2 TISSUE FIXATION 152
2 1 Formalin 152
2 2 Bouin s fixative 152
2 3 Carnoy s fixative 152
2 4 Heidenhain s Susa fixative 153
2 5 Commercial fixatives 153
3 IMPREGNATION 153
3 1 Dehydration 153
3 2 Clearing 153
3 3 Impregnation 153
4 EMBEDDING 153
5 CRYOSTAT SECTIONS 153
6 SECTIONS-STAINING 154
6 1 Preparatory steps- 154
6 2 Staining methods 154
6 3 Hematoxylin-Eosin 154
6 4 Carbohydrates (periodic acid-Schiff procedure) 155
6 5 Lipids staining 156
6 6 Nucleic acids 158
661 Feulgen and Rossbeck method 158
662 Acridine orange method 158
6 7 Detection of microorganisms 159
671 Bacteria 159
672 Mycobacteria 159
673 Fungi 160
674 Viral inclusions 161
7 STAINING OF CELLS IN WELLS AND ON COVERSLIPS 161
7 1 Hematoxylin-eosin staining 161
7 2 May-Griinwald-Giemsa staining 162
xii Contents
8 EXFOLIATIVE CYTOLOGY 163
8 1 Sampling 163
8 2 Preparation of smears 163
8 3 Fixation 163
8 4 Staining of smears 164
841 Papanicolaou method 164
9 MOUNTING 164
10 FURTHER READING 165
18 IMMUNOCYTOCHEMICAL STAINING IN ELECTRON
MICROSCOPY 167
1 INTRODUCTION 167
2 FERRITIN 167
2 1 Fixation and embedding 168
2 2 Direct method 168
2 3 Indirect method 170
2 4 Negative staining method 170
3 HORSERADISH PEROXIDASE 170
3 1 Fixation and embedding 170
3 2 Direct method 170
3 3 Indirect method 170
331 Peroxidase-anti-peroxidase complex 171
4 COLLOIDAL GOLD 171
4 1 Monodispersed colloidal gold suspensions 172
411 15 nm gold particles 172
4128 nm gold particles 172
4 2 Gold-labeled conjugates 172
421 Protein A-gold conjugates 172
422 Protein G-gold conjugates 173
423 Antibody-gold conjugates 173
4 3 Viruses and colloidal gold 174
431 Negative staining 174
432 Ultrathin sections 175
4 3 2J Fixation 175
4322 Embedding 176
4323 Immunogold procedure 176
4324 Controls 176
5 BIBLIOGRAPHY 176
19 METABOLIC AND CHEMICAL RADIOLABELING 179
1 INTRODUCTION 179
2 NUCLEIC ACIDS 181
2 1 In vivo labeling 182
2 2 Cell-free in vitro transcription 182
2 3 Enzymatic in vitro labeling 182
231 cRNA synthesis 182
232 cDNA synthesis with reverse transcriptase 183
233 DNA synthesis by nick-translation 184
234 Multiprime system 184
235 DNA probe synthesis on synthetic matrixes 185
Contents xiii
236 DNA probe synthesis using the M13 system 186
237 End-labeling 187
2371 T4 polynucleotide kinase 187
2372 Terminal transferase 187
2373 Klenow fragment 188
2374 T4 DNA polymerase 189
2375 T4 RNA ligase 189
2 4 Chemical radiolabeling 190
241 Direct radioiodination 190
242 Indirect radioiodination 191
3 PROTEINS 191
3 1 In vivo labeling 193
3 2 In vitro translation 194
321 Preparation of the S23 wheat germ extract 194
322 Microsomal membrane preparation 194
323 Translation 195
3 3 In vitro enzymatic labeling 195
331 Lactoperoxidase 195
332 Enzymobeads 196
3 4 Chemical Radiolabeling 197
341 Direct radioiodination : 197
3411 Iodine monochloride 197
3412 Chloramine-T 197
3413 Iodobeads 197
3414 Iodo-Gen 198
342 Indirect radiolabeling 198
3421 Bolton-Hunter reagent 198
3422 Wood s reagent 199
3423 Diazotized diiodosulfanilic acid 199
3424 N-hydroxysuccinimidyl-4-azidosalicilic acid
(NHS-ASA) 200
343 Tritium labeling 201
344 Labeling with metallic radionuclides 201
4 POLYSACCHARIDES 201
5 LIPIDS 202
6 FURTHER READING 203
20 IDENTIFICATION OF VIRAL PROTEINS BY
IMMUNOPRECIPITATION 205
1 INTRODUCTION 205
2 METHOD 207
3 DISCUSSION 210
4 BIBLIOGRAPHY 211
21 IDENTIFICATION OF VIRAL PROTEINS BY WESTERN
BLOTTING 213
1 INTRODUCTION ! 213
2 METHOD 214
2 1 Electrophoretic transfer 215
2 2 Western blotting 215
xiv Contents
3 DISCUSSION 217
4 BIBLIOGRAPHY 217
22 PEPTIDE MAPPING 219
1 INTRODUCTION 219
2 PROTEIN SEPARATION IN POLYACRYLAMIDE GELS 222
3 CHEMICAL CLEAVAGE 222
3 1 Cyanogen bromide 222
3 2 N-chlorosuccinimide 222
3 3 Enzymatic cleavage 224
4 DISCUSSION 224
5 BIBLIOGRAPHY 225
23 AGAROSE GEL ELECTROPHORESIS OF DNA MOLECULES 227
1 INTRODUCTION 227
1 1 Electrophoresis apparatus 231
1 2 Agarose slab gels 231
1 3 Discussion 231
2 GENOTYPING 232
2 1 Infection of cells 232
2 2 DNA preparation 234
2 3 Digestion and electrophoresis 234
2 4 Results 237
3 DISCUSSION 237
4 BIBLIOGRAPHY 239
24 EXTRACTION OF VIRAL NUCLEIC ACIDS 241
1 INTRODUCTION 241
2 EXTRACTION OF VIRAL NUCLEIC ACIDS 242
2 1 Extraction of genomes from purified virus 242
2 2 Extraction of viral mRNAs 243
221 Extraction of cellular and viral RNAs 244
222 Selection of poly(A)+ RNAs on oligo(dT)-cellulose
column 244
223 Discussion ; 245
2 3 Recombinant plasmid extraction by alkaline lysis 245
231 Plasmid extraction from a small volume of culture 246
232 Plasmid extraction from large volume of culture: plasmid
amplification 247
233 Discussion 248
3 BIBLIOGRAPHY 248
25 IN VITRO TRANSLATION 249
1 INTRODUCTION 24 9
2 IN VITRO TRANSLATION SYSTEMS 250
2 1 Reticulocyte lysate 250
2 2 Amphibian oocytes 251
3 CONCLUSION 252
4 BIBLIOGRAPHY 252
Contents xv
26 CLONING OF GENES FROM DNA VIRUSES 253
1 INTRODUCTION 253
1 1 Fragmentation of DNA 253
111 Complete digestion method 253
112 Partial digestion method 254
1 2 Choosing the cloning vector 254
121 Bacteriophage lambda and cosmid vectors 254
122 Plasmid vectors 255
2 METHOD 255
2 1 Isolation of viral DNA 257
211 Concentration of the virions 257
212 Purification of viral DNA 257
2 2 Partial digestion of genomic DNA 258
2 3 Vector linearization 259
2 4 Cohesive termini partial fill-in 260
2 5 Ligations 260
251 Control ligations 260
252 Viral DNA ligation with the vector 261
2 6 Transformation of E coli 261
261 Preparation of competent cells 262
262 Transformation 262
263 Plating of transformed cells 263
2631 Determination of cell viability 263
2632 Evaluation of the efficiency of transformation 263
2633 Plating of transformants and evaluation of
background 263
3 REFERENCES 263
27 SYNTHESIS OF COMPLEMENTARY DNA 265
1 INTRODUCTION 265
2 SYNTHESIS OF COMPLEMENTARY DNA FROM SINGLE-
STRANDED RNA 266
2 1 Synthesis of the first strand using oligo-dT primers 266
2 2 Synthesis of the first strand using small synthetic oligo
nucleotides 268
2 3 Synthesis of the second-strand cDNA 269
2 4 Digestion with SI nuclease 270
2 5 Synthesis of second-strand cDNA using RNAse-H 270
3 DISCUSSION 271
4 BIBLIOGRAPHY 271
28 SPREADING AND VISUALIZATION OF NUCLEIC ACID
MOLECULES IN ELECTRON MICROSCOPY 273
1 INTRODUCTION 273
2 MATERIAL 274
2 1 Nucleic acids 274
2 2 Glassware 274
2 3 Solutions 274
2 4 Grids 274
2 5 Cytochrome c 274
xvi Contents
3 TRANSFER TECHNIQUES 276
3 1 Protein monolayer 276
311 Macrospreading technique 276
3111 Double-stranded DNA or RNA 276
3112 Single-stranded RNA 277
312 Single-drop spreading 278
313 Microdrop diffusion 278
3 2 Non-proteinic preparative techniques 278
4 CONTRASTING THE NUCLEIC ACIDS 278
4 1 Staining with uranyl acetate 278
4 2 Shadow casting method 279
5 MOLECULAR WEIGHT DETERMINATIONS 280
6 BIBLIOGRAPHY 281
29 MOLECULAR HYBRIDIZATION 283
1 INTRODUCTION 283
2 IN SITU HYBRIDIZATION OF BACTERIAL COLONY 283
2 1 Method 284
211 Fixation and denaturation on filter papers 284
212 Fixation and denaturation on nitrocellulose filters 285
3 SOUTHERN BLOT HYBRIDIZATION 285
3 1 Fractionation of DNA 286
3 2 Preparations for transfer on filters 286
321 Nitrocellulose filters 286
322 Nylon filters 286
3 3 Fragmentation and denaturation of DNA 287
3 4 Transfer and DNA immobilization 287
341 Nitrocellulose filters 287
342 Nylon 287
4 NORTHERN BLOTTING 288
4 1 Fractionation of RNA 288
411 Glyoxal gels 288
412 Formaldehyde gels 288
413 Methylmercuric hydroxide gels 289
4 2 Preparation for transfer to filters 289
421 Nitrocellulose filters 289
422 Nylon filters 289
4 3 Transfer and RNA immobilization 289
5 HYBRIDIZATION 289
5 1 Hybridization using radioactive DNA probes 290
511 Pre-hybridization 290
512 Hybridization 290
5 2 Hybridization using oligonucleotide probes 290
521 Pre-hybridization 291
522 Hybridization 291
5 3 Hybridization using RNA probes 291
531 Pre-hybridization 291
532 Hybridization 291
5 4 Biotin-labeled probes 292
541 Pre-hybridization 292
Contents xvii
542 Hybridization 292
6 DETECTION OF HYBRIDS AND REUSE OF FILTERS 292
7 VIRUS DETECTION BY HYBRIDIZATION 294
7 1 Single-stranded RNA or DNA viral genome 294
7 2 Double-stranded RNA or DNA viral genome 294
8 CONCLUSION 294
9 BIBLIOGRAPHY 295
30 POLYMERASE CHAIN REACTION 297
T INTRODUCTION 297
2 GENERAL PRINCIPLES 297
3 SOLUTIONS 299
4 METHOD 300
4 1 RNA Amplification 300
411 cDNA synthesis by reverse-transcription 300
412 PCR amplification 300
4 2 DNA Amplification 300
4 3 Analysis of PCR products 301
4 4 Cloning of PCR products 301
4 5 Restriction endonuclease digestion 301
4 6 Ligation 302
5 BIBLIOGRAPHY 302
INDEX 303
|
| any_adam_object | 1 |
| building | Verbundindex |
| bvnumber | BV008344875 |
| callnumber-first | Q - Science |
| callnumber-label | QR385 |
| callnumber-raw | QR385 |
| callnumber-search | QR385 |
| callnumber-sort | QR 3385 |
| callnumber-subject | QR - Microbiology |
| classification_rvk | WC 5700 |
| ctrlnum | (OCoLC)28017824 (DE-599)BVBBV008344875 |
| dewey-full | 576/.64/028 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 576 - Genetics and evolution |
| dewey-raw | 576/.64/028 |
| dewey-search | 576/.64/028 |
| dewey-sort | 3576 264 228 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Book |
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| id | DE-604.BV008344875 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T17:18:41Z |
| institution | BVB |
| isbn | 0824791010 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-005513855 |
| oclc_num | 28017824 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-19 DE-BY-UBM DE-20 DE-188 DE-578 |
| owner_facet | DE-355 DE-BY-UBR DE-19 DE-BY-UBM DE-20 DE-188 DE-578 |
| physical | XVII, 309 S. Ill., graph. Darst. |
| publishDate | 1993 |
| publishDateSearch | 1993 |
| publishDateSort | 1993 |
| publisher | Dekker |
| record_format | marc |
| spelling | Methods and techniques in virology Pierre Payment and Michel Trudel ... New York u.a. Dekker 1993 XVII, 309 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Virology cabt Techniques cabt Virology Technique Virology methods Methode (DE-588)4038971-6 gnd rswk-swf Virologie (DE-588)4063597-1 gnd rswk-swf Virologie (DE-588)4063597-1 s Methode (DE-588)4038971-6 s DE-604 Payment, Pierre Sonstige oth Trudel, Michel Sonstige oth HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=005513855&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Methods and techniques in virology Virology cabt Techniques cabt Virology Technique Virology methods Methode (DE-588)4038971-6 gnd Virologie (DE-588)4063597-1 gnd |
| subject_GND | (DE-588)4038971-6 (DE-588)4063597-1 |
| title | Methods and techniques in virology |
| title_auth | Methods and techniques in virology |
| title_exact_search | Methods and techniques in virology |
| title_full | Methods and techniques in virology Pierre Payment and Michel Trudel ... |
| title_fullStr | Methods and techniques in virology Pierre Payment and Michel Trudel ... |
| title_full_unstemmed | Methods and techniques in virology Pierre Payment and Michel Trudel ... |
| title_short | Methods and techniques in virology |
| title_sort | methods and techniques in virology |
| topic | Virology cabt Techniques cabt Virology Technique Virology methods Methode (DE-588)4038971-6 gnd Virologie (DE-588)4063597-1 gnd |
| topic_facet | Virology Techniques Virology Technique Virology methods Methode Virologie |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=005513855&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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