Search

In Chapter 10, we describe two methods of complementary DNA (cDNA) synthesis and amplification. In this chapter, we will refer to these two methods of cDNA synthesis and amplification as "method A" (the technique based on classical double-stranded cDNA synthesis and subsequent adapter ligation, implemented in the Marathon kit from Clontech) and "method B" (based on the template-switching effect, implemented in the SMART cDNA synthesis kit from Clontech). These methods not only allow for amplification of representative cDNA populations from microscopic tissue samples but also provide an excellent starting point for amplification of unknown flanks of a known cDNA fragment. There are many techniques designed for this purpose (1-10); among gene hunters, they go under the generic name RACE (rapid amplification of cDNA ends). It must be remembered, though, that this name formally belongs to a particular technique introduced by Frohman et al. (8). [ABSTRACT FROM AUTHOR]

Until recently, gene arrays could only be printed on two types of supports: nylon membranes or glass slides. Nylon membrane-based arrays allow researchers to analyze hundreds of genes in a single experiment using standard laboratory equipment. However, the density of genes that can be included on] a membrane array is limited by the printing resolution. Glass arrays allow for a high printing density but can be expensive and require the use of specialized equipment. In response, BD Biosciences Clontech has developed a new type of array support, the BD Atlas™ Plastic Film, which combines the simplicity of nylon arrays with the high gene density of glass arrays. [ABSTRACT FROM AUTHOR]

Nucleic acid arrays provide a powerful methodology for studying biological systems on a genomic scale. BD Atlas™ Arrays, developed by BD Biosciences Clontech, are expression profiling products specifically designed to be accessible to all laboratories performing isotopic blot hybridization experiments. We have developed two types of readily accessible BD Atlas Arrays: nylon macroarrays, well suited for high-sensitivity expression profiling using a limited gene set, and broad-coverage plastic microarrays, for a more extensive analysis of a comprehensive set of genes. In this chapter, we describe protocols for printing and performing gene expression analysis using nylon membrane-based arrays. For a more in-depth description and protocols related to plastic film-based arrays, please refer to Chapter 3. [ABSTRACT FROM AUTHOR]