Ebook: Techniques in Molecular Biology

The last few years have seen the rapid development of new methodology in the field of molecular biology. New techniques have been regularly introduced and the sensitivity of older techniques greatly improved upon. Developments in the field of genetic engineering in particular have contributed a wide range of new techniques. The purpose of this book therefore is to introduce the reader to a selection of the more advanced analytical and preparative techniques which the editors consider to be frequently used by research workers in the field of molecular biology. In choosing techniques for this book we have obviously had to be selective, and for the sake of brevity a knowledge of certain basic biochemical techniques and terminology has been assumed. However, since many areas of molecular biology are developing at a formidable rate and constantly generating new terminology, a glossary of terms has been included. The techniques chosen for this book are essentially based on those used in a series of workshops on 'techniques in molecular biology' that have been held at The Hatfield Polytechnic in recent years. In choosing these chapters we have taken into account many useful suggestions and observations made by participants at these workshops. Each chapter aims to describe both the theory and relevant practical details for a given technique, and to identify both the potential and limitations of the technique. Each chapter is written by authors who regularly use the technique in their own laboratories.

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Content:
Front Matter....Pages 1-9
High Performance Liquid Chromatography of Proteins and Nucleic Acids....Pages 11-24
Analytical Electrophoretic Techniques in Protein Chemistry....Pages 25-48
Protein Blotting....Pages 49-61
The Production and Isolation of Peptides for Sequence Analysis....Pages 63-85
Protein and Peptide Sequence Determination....Pages 87-112
The Extraction and Fractionation of RNA....Pages 113-133
Protein Synthesis in vitro ....Pages 135-158
Labelling DNA in vitro — Nick Translation....Pages 159-166
Synthesis of cDNA for Molecular Cloning....Pages 167-183
Plasmid Isolation....Pages 185-195
The Use of Restriction Endonucleases and T4 DNA Ligase....Pages 197-219
Molecular Cloning of cDNA: Bacterial Transformation and Screening of Transformants....Pages 221-238
The Use of Minicells and Maxicells to Detect the Expression of Cloned Genes....Pages 239-255
Gel Electrophoresis of DNA....Pages 257-271
Detection of Specific DNA Sequences — The Southern Blot....Pages 273-285
The Determination of DNA Sequences....Pages 287-307
The Use of Cosmids as Cloning Vehicles....Pages 309-321
Back Matter....Pages 322-333

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Content:
Front Matter....Pages 1-9
High Performance Liquid Chromatography of Proteins and Nucleic Acids....Pages 11-24
Analytical Electrophoretic Techniques in Protein Chemistry....Pages 25-48
Protein Blotting....Pages 49-61
The Production and Isolation of Peptides for Sequence Analysis....Pages 63-85
Protein and Peptide Sequence Determination....Pages 87-112
The Extraction and Fractionation of RNA....Pages 113-133
Protein Synthesis in vitro ....Pages 135-158
Labelling DNA in vitro — Nick Translation....Pages 159-166
Synthesis of cDNA for Molecular Cloning....Pages 167-183
Plasmid Isolation....Pages 185-195
The Use of Restriction Endonucleases and T4 DNA Ligase....Pages 197-219
Molecular Cloning of cDNA: Bacterial Transformation and Screening of Transformants....Pages 221-238
The Use of Minicells and Maxicells to Detect the Expression of Cloned Genes....Pages 239-255
Gel Electrophoresis of DNA....Pages 257-271
Detection of Specific DNA Sequences — The Southern Blot....Pages 273-285
The Determination of DNA Sequences....Pages 287-307
The Use of Cosmids as Cloning Vehicles....Pages 309-321
Back Matter....Pages 322-333
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