One could be led to believe that a molecular biologist armed with a copy of 'Maniatis', or one of the 'Current Protocols' publications, would have adequate technical support to successfully accomplish most experimental procedures. In the real laboratory world, we know that even established methodology is adapting and changing at an alarming rate and that new experimental approaches are regularly appearing on the horizon. This small book fills an important niche in the market, for it aims, and I believe succeeds, in bringing the reader up to date with recent innovations in established techniques as well as introducing us to more state of the art methodology. The book contains ten chapters, all written by experts in the particular fields and interestingly, the editors have recruited over half the authors from the commercial sector. These contributions tend to bias their chapters towards products available from their particular companies, although in general they seem to have covered their subjects fairly comprehensively. Each chapter covers a review of the technique, concentrating on recent innovations and then discusses likely future trends. Most chapters end with protocols covering recent advances or more specialised approaches. Each chapter is also accompanied by an extensive list of references, in most cases concentrating on papers published in the last five years. All chapters refer to material published last year, which is a good indication that the editors and the publisher have succeeded in bringing this book to the bookshelves without undue delay.
The first chapter covers general PCR techniques and is written by a group of authors from Stratagene. In addition to covering recent advances in PCR methodology and instrumentation, the authors describe specific techniques such as cloning PCR-generated fragments and using PCR for site-directed mutagenesis. Sadly the accompanying figures are black and white copies of coloured diagrams from the company¹s catalogue and some of the detail has been lost during reproduction. A specific utilisation of PCR, thermal cycle sequencing, is described in the next chapter, which contains a generalised protocol for the technique. This is followed by a chapter devoted to methods for isolating plasmid DNA from mini-preps using silica-based resins. Whilst there are a profusion of commercial kits available, the author very rightly draws attention to the dangers of total reliance on these products and so presents a very extensive protocol utilising common laboratory reagents and equipment.
Electrophoresis is covered by three chapters, the first by Branko Kozulic, who provides a very readable account of recent theories which attempt to explain electrophoretic phenomena, including his own Œdoor-corridor¹ model. He also provides a tantalizing glimpse into the world of new gel matrices and intercalating dyes. The second chapter is devoted to pulsed field gel electrophoresis (PFGE) in which the authors review the various aspects of the technique and provide protocols for the preparation of high molecular weight DNA from soya bean leaves and provide physical mapping data from PFGE combined with two dimensional electrophoresis. The other chapter describes capillary electrophoresis (CE) as applied to the isoelectric focusing of proteins and provides an extensive protocol and a troubleshooting chart.
A chapter on subtractive hybridisation describes the use of commercially available multipurpose cloning vectors to perform cDNA subtractive hybridisation between biotinylated RNA and single stranded DNA. The unhybridised product is purified by streptavidin and used for transformation. This technique should appeal to researchers involved in gene expression and developmental studies.
The widespread use of PCR in molecular biology has required the simultaneous development of reliable methods for the production of oligo primers. A chapter describes recent developments in the related field of oligoribonucleotide synthesis. The demand for synthesized RNA is likely to increase as interest in antisense RNA and the possible use of ribozymes in gene therapy intensifies.
Finally, there are two interesting chapters on instrumentation. One describes state of the art devices for automated DNA hybridization and detection and the other is devoted to a relatively new technique called matrix assisted laser desorption ionization mass spectrometry (MALDI). The authors speculate that MALDI will, in the not too distant future, replace gel electrophoresis in the analysis of DNA sequencing reactions.
This modestly priced book provides the molecular biologist with a wealth of current information on a wide variety of essential techniques. I look forward to the publication of volume 2 in this series, later this year.
http://www.horizonpress.com/hsp/revs/revs1mb.html
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