In situ hybridization: a practical approach
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Oxford [u.a.]
Oxford Univ. Pr.
1998
|
| Edition: | 2. ed. |
| Series: | The practical approach series
196 |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | XVIII, 224 S. Ill. |
| ISBN: | 0199636591 0199636583 |
Staff View
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| 245 | 1 | 0 | |a In situ hybridization |b a practical approach |c ed. by D. G. Wilkinson |
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Record in the Search Index
| _version_ | 1804126975445434368 |
|---|---|
| adam_text | In Situ
Hybridization
A Practical Approach
Second Edition
Edited by
D G WILKINSON
NIMR, London
Oxford New York Tokyo
OXFORD UNIVERSITY PRESS
Contents
List of contributors
Abbreviations
1 The theory and practice of in situ
hybridization
David G Wilkinson
1 Introduction
2 Preparation ot probe
Type of probe and method of synthesis
Length of probe
Label
3 Preparation of tissue
Fixation
Embedding and sectioning of tissue
Subbing of slides
4 Pre-treatments of tissue
5 Hybridization and washing
Optimal conditions for hybridization
Post-hybridization washing
6 Visualization of signal
Radioactive probes
Hapten labelled probes
Double staining techniques
7 Controls and troubleshooting
8 Photography
Acknowledgements
References
2 Oligonucleotide probes for in situ
hybridization
Marcus Rattray and Gregory J Michael
1 Introduction
Contents
2 Design of oligonucleotide probes 23
Theoretical considerations 23
Design in practice 27
3 Oligonucleotide probe synthesis 29
Manufacturing oligonucleotides 29
Oligonucleotide purification 29
Quantification of oligonucleotide 32
4 Oligonucleotide labelling 33
Choice of labelling method 33
3 end-labelling with radiolabelled nucleotides 34
Non-radioactive 3 end-labelling 38
5 In situ hybridization with oligonucleotides 39
Tissue preparation 39
Hybridization and post-hybridization washes 45
Combined in situ hybridization with immunocytochemistry 51
Controls for in situ hybridization S3
6 Detection of hybrids and quantification 55
Detection of radiolabelled probes 55
Imaging and analysis of sections hybridized with radiolabelled probes 61
Image analysis using non-radioactive probes 64
7 Stock solutions and precautions against RNase contamination 64
References 66
3 Detection of mRNA in tissue sections with
radiolabelled riboprobes 69
Antonio Simeone
1 Introduction 69
2 Recovery and fixation of embryos 69
3 Wax embedding, orientation, and sectioning - 71
4 Probe preparation 75
5 Pre-treatment and hybridization 76
6 Post-hybridization washes 78
7 Autoradiography 79
8 Developing, staining, and photography 80
9 In situ hybridization on cryosections 82
10 Useful molecular markers to define gene expression patterns 83
Acknowledgements 84
References 84
Contents
4 In situ hybridization of mRNA with hapten
labelled probes 87
Qiling Xu and David G Wilkinson
1 Introduction 87
2 When to hybridize to sections or whole mounts 87
3 General strategy 88
4 Whole mount in situ hybridization 89
Preparation of labelled RNA probe 90
Fixation and pre-treatment of embryos 92
Hybridization, washing, and detection of probe 95
5 In situ hybridization to tissue sections or cultured cells 99
General precautions 99
Preparation of tissue sections or cultured cells on slides 100
Pre-treatmentS; hybridization, washing, and detection of probe 101
6 Photography 104
Acknowledgements 106
References 106
5 Two colour in situ hybridization 107
T Jowett
1 Methods of localization of multiple transcripts 107
Two colour in situ hybridization with chromogenic substrates for
alkaline phosphatase and horse-radish peroxidase 110
Two colour in situ hybridization with chromogenic substrates for
alkaline phosphatase 113
Double fluorescent in situ hybridization 117
2- Methods for simultaneous in situ localization of transcripts
and tissue antigens 121
Indirect immunolocalization of a tissue antigen 122
Immunolocalization by the peroxidase anti-peroxidase (PAP) method 123
Immunolocalization by the ABC method 124
References 126
6 Electron microscope in situ hybridization: the
non-isotopic post-embedding procedure 127
Francine Puvion-Dutilleul
1 Introduction 127
xi
Contents
2 Preparation of biological material for ultrathin sectioning 128
Fixation 128
Dehydration and Lowicryl embedding 129
Supports for ultrathin sections 130
Sectioning and recovering ultrathin sections 131
3 Probes 131
Generalities 131
Probe choice 131
4 Hybridization 133
Generalities 133
Hybridization solution 133
Heat treatment of the hybridization solutions 134
Formation of hybrids on ultrathin sections 134
Post-hybridization steps 136
Hybridization protocol 137
5 Specific preparations of target molecules prior to
hybridization 138
Why and how does one improve the accessibility of the probe to
the target? 138
Why and how to perform nuclease digestion(s) prior to hybridization 140
6 Controls of the specificity of the detection 141
7 Routine protocols for specific detection of defined nucleic
acid sequences and applications 142
Specific detection of DNA sequences by using biotinylated double-
stranded DNA probes 142
Specific detection of RNA sequences by using biotinylated probes 148
8 Double detections 155
Simultaneous detection of two distinct nucleic acid sequences 155
Simultaneous detection of a nucleic acid sequence and an antigen 156
9 Conclusions 156
Acknowledgements 157
References 157
7 Detection of genomic sequences by
fluorescence in situ hybridization to
chromosomes i6i
Lyndal Kearney
1 Introduction 161
2 FISH: practical considerations 162
Preparation of metaphase chromosomes 162
xii
Contents
Probes for FISH
Pre-treatment of chromosomal DNA
Competitive in situ suppression hybridization
Detection of bound, labelled probe
Visualization of signal
3 Troubleshooting
4 Applications
Gene mapping
Identification of genetic material of unknown origin
Comparative genomic hybridization (CGH)
Breakpoint identification
Interphase analysis
Comparative mapping
Gene expression, structure, and function
References
8 Gene expression databases 190
Duncan Davidson, Richard Baldock, Jonathan Bard,
Matthew Kaufman, Joel E Richardson, Janan T Eppig,
and Martin Ringwald
1 Introduction 190
2 The database approach: combining information in a
common format 190
Descriptions of embryos: information formats for developmental
biology 190
Molecular anatomy: a new description of the embryo 191
Using gene expression data to build molecular pathways 191
Linking pathways with processes at the cell and tissue levels 192
Problems with the database approach 192
3 Outline survey of gene expression databases 193
Caenorhabditis 194
Drosophila 194
Zebrafish 195
Xenopus 196
Mouse 196
Human 204
Specialist data 205
4 Using gene expression databases 205
Practical considerations in preparing to enter data into a database 205
Querying the database 211
5 The future of gene expression databases 212
Between-species links 213
xiii
Contents
Acknowledgements 213
References 214
Appendix 215
Index 221
XIV
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| spelling | In situ hybridization a practical approach ed. by D. G. Wilkinson 2. ed. Oxford [u.a.] Oxford Univ. Pr. 1998 XVIII, 224 S. Ill. txt rdacontent n rdamedia nc rdacarrier The practical approach series 196 In Situ Hybridization Laboratory Manuals In situ hybridization Laboratory manuals In-situ-Hybridisierung (DE-588)4215968-4 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf DNS (DE-588)4070512-2 gnd rswk-swf DNS (DE-588)4070512-2 s In-situ-Hybridisierung (DE-588)4215968-4 s Methode (DE-588)4038971-6 s DE-604 Wilkinson, David G. Sonstige oth The practical approach series 196 (DE-604)BV007921321 196 HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008373168&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | In situ hybridization a practical approach The practical approach series In Situ Hybridization Laboratory Manuals In situ hybridization Laboratory manuals In-situ-Hybridisierung (DE-588)4215968-4 gnd Methode (DE-588)4038971-6 gnd DNS (DE-588)4070512-2 gnd |
| subject_GND | (DE-588)4215968-4 (DE-588)4038971-6 (DE-588)4070512-2 |
| title | In situ hybridization a practical approach |
| title_auth | In situ hybridization a practical approach |
| title_exact_search | In situ hybridization a practical approach |
| title_full | In situ hybridization a practical approach ed. by D. G. Wilkinson |
| title_fullStr | In situ hybridization a practical approach ed. by D. G. Wilkinson |
| title_full_unstemmed | In situ hybridization a practical approach ed. by D. G. Wilkinson |
| title_short | In situ hybridization |
| title_sort | in situ hybridization a practical approach |
| title_sub | a practical approach |
| topic | In Situ Hybridization Laboratory Manuals In situ hybridization Laboratory manuals In-situ-Hybridisierung (DE-588)4215968-4 gnd Methode (DE-588)4038971-6 gnd DNS (DE-588)4070512-2 gnd |
| topic_facet | In Situ Hybridization Laboratory Manuals In situ hybridization Laboratory manuals In-situ-Hybridisierung Methode DNS |
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