Basic Cloning Procedures

✍ Scribed by Juris Steinbergs, Aleksander Tsimanis (auth.), Professor Dr. Valdis Berzins (eds.)

• Publisher: Springer-Verlag Berlin Heidelberg
• Year: 1998
• Tongue: English
• Leaves: 174
• Series: Springer Lab Manual
• Edition: 1
• Category: Library

✦ Synopsis

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.

✦ Table of Contents

Front Matter....Pages i-vii
cDNA Synthesis and Cloning....Pages 1-21
DNA Sequencing....Pages 22-42
RNA In Vitro Synthesis by Phage T7 DNA-Dependent RNA Polymerase....Pages 43-52
PCR-Based Site-Specific Mutagenesis....Pages 53-67
Analysis of Specific Protein-DNA Interactions....Pages 68-103
Gel Electrophoresis, Transfer and Hybridization of Nucleic Acids....Pages 104-134
Immunological Methods for Analysis of Recombinant Proteins....Pages 135-144
Protein Synthesis in Cell-Free Systems....Pages 145-163
Back Matter....Pages 165-169

✦ Subjects

Biochemistry, general; Cell Biology