Practical Guide to Molecular Cloning

About this book

This new edition of the standard laboratory manual is intended to make molecular cloning faster and easier. It presents techniques tested at the Curie Institute and at other leading laboratories, and lists all commercially available enzymes, vectors, linkers, and other basic products for ready reference. A detailed explanation of protocols is offered, allowing the isolation, cloning, and expression of genes from living species. As well as presenting up-to-date techniques on sequencing, in vitro expression of cloned gene and use of computers for study of nucleic acids, it shows how to isolate DNA-protein complexes and new methods for mutagenesis of cloned genes.

Contents

Laboratory Equipment Needed for Molecular Cloning; A Few Words about Safety; Basic Methods in Molecular Biology; Useful Modifying Enzymes for Molecular Cloning; Restriction Endonucleases; Vectors for Molecular Cloning; Purification and Characterization of Vector and Passenger DNA; Digestion of DNA with Restriction Endonucleases; Separation of DNA Fragments by Electrophoresis; Ordering the Restriction DNA Fragments in Physical Maps; Purification of DNA Fragments; Modification of DNA Fragments with Cohesive Termini; Ligation; Propagation of Recombinant DNA Molecules; Characterization of Recombinant Clones; Preparation of Genomic Libraries; Purification and Characterization of RNA Species; Cloning of cDNA Species; Sequencing of DNA; RNA Sequencing; Use of Computerized Programs in the Treatment of DNA Sequence Information; Localized Mutagenesis; Methods for Studying DNA-Protein Interactions; Expression of Cloned DNA Sequences In Vitro or In Procaryotic and Eucaryotic Cells; Appendixes.

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