Differential Display Methods and Protocols

Based on a new mathematical model of differential display (DD), this updated second edition takes advantage of automation, as well as digital data acquisition and analysis. The authors explain and highlight all the recent methodological refinements, including automated liquid handling of hundreds of DD PCR reaction setups combined with capillary electrophoresis, a prototype computer program to automatically allow positive band identification from a fluorescence differential display (FD) image, and restriction fragment-based DD screenings. Other improvements discussed are the combining of DD and DNA microarrays by reducing the complexity of cDNA probes while increasing the sensitivity of detection, and a DD approach to detect prokaryotic mRNA expression. The authors also demonstrate the power of DD technology with a collection of outstanding examples of DD applications and detailed experimental procedures. The elegant studies described here have led to the discovery of many important genes involved in viral infection, Prion disease, cancer, ovulation, circadian clock, floral color, transcription repression gene silencing, mRNA polymorphism, and protein-RNA interaction.Carrying on the high standards of the first edition of Differential Display Methods, Peng Liang et al. have based their second edition on a new mathematical model of differential display (DD) that takes advantage of automation, as well as digital data acquisition and analysis. These well-versed authors explain and highlight all the recent methodological refinements, including automated liquid handling of hundreds of DD PCR reaction setups combined with capillary electrophoresis, a prototype computer program to automatically allow positive band identification from a fluorescence differential display image, and restriction fragment-based DD screenings that can link any cDNA fragment directly to a given gene once the sequence information of all transcripts becomes available. Other improvements discussed are combining DD and DNA microarrays by reducing the complexity of cDNA probes while increasing the sensitivity of detection, and a DD approach to detect prokaryotic mRNA expression. The authors also demonstrate the power of DD technology with a collection of outstanding examples of DD applications and detailed experimental procedures. The elegant studies described here have led to the discovery of many important genes involved in viral infection, Prion disease, cancer, ovulation, circadian clock, floral color, transcription repression gene silencing, mRNA polymorphism, and protein-RNA interaction. §State-of-the-art and highly practical, Differential Display Methods, Second Edition offers gene hunters the possibility of genome-wide comprehensive DD screening, as well as a proven road map for any successful "gene fishing" expedition.