Gene Cloning and Analysis: Current Innovations Book Reviews

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Review by
CAB International

April 1997

This multiauthor book has 13 chapters and a 12 page index, and focuses on newly emerging technologies that facilitate the isolation and characterization of genes. The majority of methods are applicable to any biological system. Subject coverage includes the yeast 2-hybrid system, peptide nucleic acid (PNA) for nucleic acid capture, polymerase chain reaction (PCR) for isolation of flanking DNA, RNA ligase-mediated random amplified cDNA ends (RACE), site directed mutagenesis, PCR-ligation-PCR-mutagenesis, universal TA cloning of PCR products, long PCR, simple sequence conformation polymorphism (SSCP) analysis, cloning by protein ligand interaction, and gene expression fingerprinting. Each chapter contains protocols with notes and troubleshooting tips, and concise background information, frequently with a review of similar methods followed by an assessment of future innovations.

Review by
Irina R. Tsaneva University College, London.

SGM Quarterly, August 1997

Amongst the avalanche of commercial kits and technical bulletins, I found this book pleasantly refreshing and stimulating. It presents a selection of newly emerged technologies of the higher scientific stan- dard with an emphasis on innovation and creativity. Some of the methods described may become routine in many labs and others would stimulate new developments. I would therefore welcome this new addi- tion to the numerous published protocols on gene cloning and analysis. The target audience of this book is most likely to be the seasoned investigator. 'Maniatis' proficiency would be essential as many of the described techniques go well beyond the kitchen of everyday molecular biology. Some would require an effort to set up, but the initial investment should be repaid with interest.
The book is well suited for personal purchase in specialized groups. Research-conscious institutions would be well advised to acquire this book for the infrequent user.

Review by
Mike Starkey UK Human Genome Mapping Project, Resource Centre, Hinxton, Cambridge.

Heredity, August 1997

The major theme of this book is refinements to existing technologies and recently innovated technologies for the isolation and partial characterization of genes. The topics are particularly pertinent to the functional analysis phase of the human genome project given the identification of large numbers of cDNAs via Expressed Sequence Tag studies. A restriction endonuclease-based differential display approach for identifying cDNAs of interest is detailed in chapter 3, whilst the isolation of full length cDNAs is described by chapters 6 and 7. Elsewhere is considered the characterization of cDNA-encoded proteins on the basis of protein-protein/ligand interaction (chapters 1 and 2), codon mutagenesis to study the rela tionship between protein structure and function (chapter 8), and the alteration of protein function by gene replace ment (chapter 10).
The remainder of the articles is loosely divided into techniques of perhaps particular relevance to diagnostics, and procedures that are applicable to molecular biologists working in a spectrum of fields. Peptide nucleic acid based purification techniques (chapter 4) are suited to the sequence-specific purification of DNA templates from clinical material, whilst PCR-SSCP analysis (chapter 13) remains a valuable tool for the detection of sequence variation in DNA samples. Of universal interest, are TA cloning (chapter 11), a PCR-based method for isolating genomic DNA flanking a known sequence (chapter 5), and a method combining restriction digestion with 'long PCR' to effect selective amplification of one of a group of homologous sequences (chapter 12).
Volumes of this genre are normally a joy for the bench scientist because they elaborate on the nuances of experimental procedures and fill in the unwritten gaps so often present in scientific papers (indeed, there are few things more satisfying than being able properly to apply some thing that one has read to good effect in a short period of time). With few exceptions, this book fulfils the role of a good laboratory companion. Furthermore, the information is often usefully presented in combination with a review of variants of the procedure discussed. Chapter format is consistent throughout; a review of alternative approaches to a given technology is followed by a description of a specific procedure, preceding a section on troubleshooting and useful tips. The level of knowledge required for effective use varies from chapter to chapter; in the case of the more involved chapters, the protocols are not as complete as in roughly equivalent texts, and referral to other references is required. For this reason the book is probably best targeted at the reasonably well informed. However, in essence the book is well compiled and a jolly good read for the practising molecular biologist.