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.

Inhalt

Juris Steinbergs, Aleksander Tsimanis: cDNA Synthesis and Cloning.- Eriks Jankevics: DNA Sequencing. - Elita Avota, Normunds Licis: RNA in Vitro Synthesis by Phage T7 DNA-Dependent RNA Polymerase.- Janis Klovins, Valdis Berzins: PCR-Based Site-Specific Mutagenesis.- Indrikis Muiznieks, Nils Rostoks: Analysis of Specific Protein--DNA Interactions.- Illar Pata, Erkki Truve: Gel Electrophoresis, Transfer and Hybridization of Nucleic Acids.- Irina Sominskaya, Kaspars Tars: Immunological Methods for Analysis of Recombinant Proteins.- Tiina Tamm, Erkki Truve: Protein Synthesis in Cell-Free Systems.