This manual introduces the reader to bacic methods used in the isolation, cloning and analysis of genetic material. The protocols include RT-PCR amplification, gene cloning, hybridization analysis and sequencing of nucleic acids, PCR-based site-specific mutagenesis, analysis of protein DNA-specific interaction, cell-free protein synthesis and product electrophoretic and immunological analysis. Each protocol includes short background information, a detailed description of the necessary materials, step-by-step procedures, a troubleshooting guide and useful practical hints.

1 cDNA Synthesis and Cloning

2 DNA Sequencing
3 RNA In Vitro Synthesis by Phage T7 DNA-Dependent RNA Polymerase
4 PCR-Based Site-Specific Mutagenesis
5 Analysis of Specific Protein-DNA Interactions
6 Gel Electrophoresis, Transfer and Hybridization of Nucleic Acids
7 Immunological Methods for Analysis of Recombinant Proteins
8 Protein Synthesis in Cell-Free Systems.