Microarrays and Microplates: Applications in Biomedical Sciences

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ISBN: 9781859960745 | 185996074X
Cover: Nonspecific Binding
Copyright: 6/25/2003

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Microarrays and Microplatestitle looks at the new microarray and microplate-based technologies which facilitate large-scale analysis of DNA sequence variants, mRNA molecules and proteins. The book provides a review of the various methodologies being used to identify genetic variants and gene regulation and guides readers through both the application of these methodologies and experimental procedures.

Contributors	p. vii
Abbreviations	p. viii
Preface	p. x
Instrumentation for Genomics and Proteomics	p. 1
Introduction	p. 1
History of the microplate	p. 1
Microplate-handling instrumentation	p. 2
Applications in genomics	p. 8
Applications in proteomics	p. 16
Conclusion	p. 18
Arraying of PCR products onto nylon membranes	p. 20
Processing colony arrays	p. 21
Expression Microarrays	p. 23
Introduction	p. 23
Microarrays in overview	p. 23
Array fabrication	p. 26
Preparation of labelled probes	p. 29
Microarray hybridization	p. 29
Generation and analysis of expression data	p. 30
The need for databases	p. 33
Validation of gene expression changes	p. 33
Summary and future prospects	p. 34
PCR amplification of cDNAs	p. 37
PCR product purification by precipitation	p. 38
Post-processing of glass slides	p. 39
Fluorescent probe labelling	p. 40
Pre-hybridization	p. 42
Microarray hybridization and washing	p. 43
Tissue Microarrays: A High-Throughput Technology for Translational Research	p. 45
Introduction	p. 45
Tissue microarray construction	p. 46
Staining of tissue microarrays	p. 49
Tissue microarray data and image management	p. 51
Applications in translational research	p. 54
Validation of tissue microarrays	p. 57
Future developments	p. 58
Protein Chips and Microarrays	p. 61
Introduction	p. 61
Microarray technology	p. 64
Versatility of protein microchips	p. 77
Concluding remarks	p. 85
Single Nucleotide Polymorphism Genotyping Using Microarrays	p. 89
Introduction	p. 89
DNA microarrays for sequence analysis	p. 89
Arrays for resequencing and SNP-scoring	p. 92
Summary and future directions	p. 101
Multiplexed allele-specific extension coupled with tag-arrays	p. 109
Microplate-Based mRNA Analysis	p. 115
Introduction	p. 115
Microplate-based RNA purification	p. 116
Microplate-based PCR	p. 119
Microplate-based non-RT-PCR assays	p. 123
Application of mRNA analysis to drug discovery project	p. 124
Application of mRNA analysis to clinical diagnostics	p. 125
A Generalized 96-Well ELISA-Based Assay for Quantitative and Qualitative Monitoring of Cellular Events	p. 131
Introduction	p. 131
The 96-well in situ ELISA assay	p. 133
Technical considerations	p. 134
Suggested statistical treatment	p. 137
Additional applications of the assay	p. 140
Conclusion	p. 140
The 96-well in situ ELISA assay	p. 142
Electrophoresis in Microplate Formats
Introduction	p. 145
MADGE, 96-well formats	p. 148
192-Well, 384-well, 768-well MADGE, ARMS and SNP typing	p. 152
96-Well high-resolution MADGE	p. 155
Temporal thermal ramp MADGE electrophoresis (Melt-MADGE)	p. 156
Software applications (applies to all MADGE approaches)	p. 160
The future	p. 162
384-Well MADGE	p. 165
Appendix	p. 171
Index	p. 173
Table of Contents provided by Rittenhouse. All Rights Reserved.

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Microarrays and Microplates: Applications in Biomedical Sciences

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