Overview of Applications Formats

Abstract

The detection of nucleic acids by nonradioactively labeled probes can be performed either with immobilized analytes on blots, by in situ approaches or in solution. The table shows an overview of important application formats including cross reference to the formats described in Chaps. 19–22. With blot and in situ formats, the presence or absence of a particular sequence is recorded, whereas detection in solution allows quantitative measurements. For detection of nucleic acids, a number of blot formats have been established: dot blot, slot blot, Southern blot (DNA analytes), northern blot (RNA analytes), southwestern blot (protein-binding DNA sequences), and genomic blot (analysis of whole genomes). DNA sequencing on blots have also been developed (Richterich et al., 1989; Höltke et al., 1992). A variety of formats have been described for detection oc nucleic acids in situ: colony hybridization (bacterial colonies), plaque hybridization (phage plaques), in situ hybridizations with isolated metaphase chromosomes, tissue sections, biopsies, fixed cells, or whole organisms such as Drosophila embryos. Both proteins and glycoproteins are most often analyzed in western blots. For review of the alternative formats see Matthews and Kricka, 1988; Wilchek and Bayer, 1988; Kessler, 1991; Kricka, 1992.