Hybridization with Nucleic Acid Probes, Part II
Part II. Probe Labeling and Hybridization TechniquesBy
- P. Tijssen, Université de Quebec, Institut Armand Frappier, Villle de Laval, Quebec, Canada
Recent breakthroughs in recombinant DNA technology and the availability of sophisticated equipment accessible to almost any laboratory, have contributed to the development and perfection of powerful hybridization tools. Recently, nucleic acid hybridization has not only become a cornerstone in molecular biology research but also a powerful supplement to other diagnostic tools.These diagnostic methods are set out in a logical and clear two-part volume in this now famous Laboratory Techniques series. The volume is divided into theory and preparation (Part I), and probe labelling and hybridization techniques (Part II). Both parts are worthy additions to this series, designed for easy access of information on the laboratory bench.
Laboratory Techniques in Biochemistry and Molecular Biology
Published: October 1993
Excluding sub-sections. Full contents list available on request.
Chapters 1-6 are Part I of this title.
Chapter 7. Labeling of probes and their detection. 7.1. Choice of label and labeling method. 7.2. Radioisotopic labeling and detection of nucleic acids. 7.3. Nonradioactive primary labels. 7.4. Recognition systems. 7.5. Overview of enzymatic incorporation of labels into probes. 7.6. Uniform incorporation of labels in nucleic acids. 7.7. End-labeling of probes. 7.8. Chemical modification of probes to introduce labels. 7.9 Universal probes. 7.10. Biological probes. 8. Mixed-phase and solution hybridization formats. 8.1 Materials and solutions. 8.2. Slot/dot blot hybridization on membranes. 8.3. (Semi)-solution hybridization. 9. Hybridization after electrophoretic fractionation of nucleic acids. 9.1. Electrophoretic procedures. 9.2. Transfer procedures. 9.3. Hybridization in the gel or before electrophoresis. 10. Colony and plaque lift hybridization. 10.1. Colony and plaque hybridization: different approaches. 10.2. Colony hybridization. 10.3. Plaque hybridization. 10.4. Hybridization to colony or plaque nucleic acids. 10.5. Selection, packing and purification of clones. 11. In situ hybridization. 11.1. Theoretical considerations. 11.2. Cytological procedures. 11.3. Hybridization procedures. 11.4. Emerging in situ hybridization techniques. 12. Selected applications of hybridization. 12.1. Aims of subtractive or suppression hybridization. 12.2. Hybrid selection of mRNA. 12.3. S1 analysis of mRNA with DNA probes. 12.4. RNase protection assays. 12.5. Triple helices. 12.6. Current developments and prospects. References and Index (Parts I and II).