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Doody's Review ServiceReviewer: Eugene A Davidson, PhD (Georgetown University School of Medicine)
Description: This is the second edition (first edition published in 1997) of a compilation of protocols for investigators working with differential display. Both theoretical and practical aspects are addressed; several practical applications are included.
Purpose: The goal is to provide a set of protocols for investigators wishing to use differential display methods for monitoring gene expression. There are competing methods, notably array technologies, and the differences/advantages are not discussed in detail. The goal is worthwhile, but overall not well achieved.
Audience: This is essentially a practical laboratory manual and as such is intended for the senior graduate student/research fellow/investigator. There is sufficient detail to allow most of the experiments to be executed. The editors have assembled a diverse and capable group of contributors.
Features: Given the current knowledge of the genome and the ability to measure gene expression in multiple ways, it is to be expected that investigators will want to follow the "performance" of one or more genes during a variety of cellular events. These include differentiation/development, malignant transformation, sequelae of infectious processes, etc. Although the use of microarrays is very popular, these are high cost items and not available to all laboratories. Thus, alternative approaches are desirable. This book offers a series of protocols aimed at providing the tools necessary to use differential display. In principle, messenger RNA can be prepared from the tissue of interest, a cDNA copy made and then specific interrogation made of that population by PCR or other techniques. The initial chapter provides a theoretical discussion of global gene display addressing issues such as coverage of the chromosome when targets are not explicit. Subsequent chapters are laboratory oriented and include methods for automation, use of ordered displays, and PCR methods designed for this approach. The final section includes several explicit studies such as detection of genes expressed in prion disease, ovulation, and those related to p53. Each chapter has a short reference list to complement the detailed instructions. As a second edition, there is some new material. However, some sections are not very successful. Thus, the discussion of one new gene possibly identified in prion disease belongs in the primary literature awaiting confirmation. The automated procedure described is very challenging experimentally and, as with other chapters, insufficiently describes expected pitfalls and experimental snags. Investigators needing to use differential display will find useful, albeit limited, material.
Assessment: As a second edition, this book does not add very much. It is particularly important for a book of laboratory protocols to include caveats else the inexperienced investigator may be left at sea. At the same time, the experienced laboratorian is aware of the issues. The book seems cobbled together rather than tightly organized.