Genetic manipulation is no longer the province of the specialized researcher. It is finding widespread application in all fields of medicine and biology. Nevertheless, application of these relatively new techniques to new areas of research is often fraught with unexpected problems and difficulties. Based on the Society for Applied Bacteriology's Autumn 1989 Conference, this unique volume covers a wide and very up-to-date range of techniques used in genetic engineering. These include the isolation and analysis of DNA and RNA from cells and tissues, the selection and use of phage and plasmic vectors for cloning DNA, the cloning procedures, the production and screening of genomic libraries, the production and use of DNA probes, the polymerase chain reaction and the synthesis of 'designer' genes. This volume contains many examples of the applications of the above and other techniques for genetic manipulation, to subjects as diverse as plant pathology, forensic science, bacterial taxonomy, cardiac research, diagnostic microbiology, food hygiene and sewage treatment.
Extraction, purification and assay of DNA; Restriction mapping of DNA; The construction and use of cloning vectors; Adaptor based cDNA cloning by the phage vectors; Plasmid profiling and DNA/DNA hybridization for distinguishing between mesophilic aeromonas bacteria; Preparation and screening of genomic libraries; DNA probes for detection and identification of bacteria; Extraction and purification of eukaryotic mRNA; cDNA cloning; Primer extension sequencing of RNA viruses; Forensic applications of DNA profiling; Application of nucleic acid probes to the identification of bacterial enteric pathogens; Quantification of collagen mRNA levels in mammalian cells; Detection of food-borne bacterial pathogens by a colorimetric DNA hybridization method; Use of a DNA probe for a viroid in plant pathology; Analysis of restriction fragment length polymorphisms in the study of bacteria; The use of DNA probes to detect 'selected bacteria' after their introduction into the waste water treatment plant; The use of genetic manipulation to study bacterial pathogens; RNA detection methods in basic cardiac research; The polymerase chain reaction and its application to basic research in molecular biology; The effect of nutrient limitation on the stability and expression of recombinant plasmids; Large scale production of proteins from recombinant DNA in Escherichia coli; Stability and copy number of yeast cloning vectors; Designer genes - Design and synthesis illustrated for the gene coding for human macrophage colony stimulating factor; Monitoring safety in process biotechnology; Patenting in biotechnology