Experiments in Molecular Biology

Research in the field of molecular biology has progressed at a fascinating rate in recent years. Much of this progress results from the development of new laboratory techniques that allow very precise fractionation and analysis of nucleic acids and proteins, as well as the construction of recom­ binant DNA molecules that can then be cloned and expressed in host cells. Progress has been so rapid that there has been a shortfall in the training of appropriately qualified staff. Many existing laboratory workers require retraining, and many educational institutions have had difficulty incor­ porating the new molecular biology techniques into their teaching programs. Although there are several manuals currently available that describe laboratory techniques in molecular biology, they are principally written for the indivi­ dual research worker and are not intended for use in the design of practical classes for students. The aim of this book is to provide just such a series of pro­ tocols for the teaching of practical molecular biology. The idea arose following the success of several Workshops in Molecular Biology, organized and taught by staff in the Biol­ ogy Department of the Hatfield Polytechnic. Gradually, the protocols used in the workshops have been incorporated into the Hatfield undergraduate and postgraduate teaching pro­ grams and have now been collected together to form a book.

Contenu

DNA Isolation and Transformation in Escherichia coli.- Isolation of Plasmid DNA.- Restriction Endonuclease Digestion and Agarose Gel Electrophoresis of DNA.- Restriction Site Mapping.- Southern Blotting.- Preparative Electrophoresis.- DNA Ligation and Escherichia coli Transformation.- Detection of Plasmid by Colony Hybridization.- Microisolation and Microanalysis of Plasmid DNA.- Isopycnic Centrifugation of DNA.- Purification of Plant Ribosomal DNA.- The Extraction and Affinity Chromatography of Messenger RNA.- Agarose Gel Electrophoresis of RNA.- In Vitro Protein Synthesis.- SDS Polyacrylamide Gel Electrophoresis of Proteins.- Isoelectric Focusing (IEF) of Proteins in Thin-Layer Polyacrylamide Gels.- Protein (Western) Blotting with Immunodetection.- Thin-Layer Gel Filtration.- The Dansyl-Edman Method for Peptide Sequencing.- The Double Immunodiffusion Technique.- Immunoelectrophoretic Techniques.- The Time Course of ?-Galactosidase Induction in Escherichia coli.- Experiments on Enzyme Induction in the Unicellular Green Alga Chlorella fusca.- Yeast Protoplast Fusion.- The Ames Test.