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A practical guide to molecular cloning
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Book reviews A Practical Guide to Molecular Clo,fing, by B. Perbal. 2nd edn. John Wiley & Sons, Chichester, 1988, 812 pp., £ 32.50 Introduction to Practical Molecular Biology, by P.D. Darbre. John Wiley & Sons, Chichester, 1988, 117 pp., £ 9.95 Everybody aware of practical molecular genetics knows the blue cover of the 'Maniatis', and it is not a rare event that it has been stolen from a shelf... This may explain why this practical textbook has had many clones as did the IBM PC microcomputers. A Practical Guide to Molecular Cloning by B. Perbai (the second edition, already!) is such a clone, but different enough from its reference to be worth buying in addition to the holy book. Safety recommendations are present at the start of this big guide (chapter 2) and this is certainly a good starting point. Then follow all standard techniques of molecular biology: gel electrophoresis and filtration, ultracentrifugation, ion-exchange chromatography, autoradiography, DNA precipitation. Not much is said about/3-galactosidase or about affinity chromatography (for instance using this enzyme as a carrier protein); it is our hope that this hole will be filled in a future edition! Restriction and modifica6on are extensively developed, useful enzymes (and also others) are presented with their target sites, a lengthy table presents a usefnl nattprn nf huhrirl c~n,,~ne,~c rpcnltlna f r n r , ~ !igation generated after digestion with different restriction enzymes. Unfortunately, however, the understandable wish to be complete results in such a large table that it becomes practically useless. A table such as the one given in the well known Biolabs catalogue would be a significant improvement in that matter. At least isoschizomers should have been grouped (why separate BamHl and BstI?). But rather than go through each chapter (the book is over 800 pages long, including protocols), let's go through the protocols. The material is E. coli K12 or B, in general, with appropriate phage, cosmid or plasmid vectors. DNA preparation is therefore of major importance. A rapid procedure described for plasmid preparation involves, all standard ingredients (SDS, NaOH, and.., lysosyme, which could be omitted without any damage to the yield and purity), but adds several steps (dialysis on Millipore filters) which I have never personally used and might, perhaps, increase the digestion efficiency of some enzymes. A thorough analysis of restriction enzymes activity as a function of salt concentration will certainly be useful to many: it is always disappointing to obtain partial digestions on a sample containing only preciously little DNA. Another chapter (13) is devoted to an ever surprising step in molecular cloning: iigation. In this chapter we
are presented with theory (which I have not checked, but which seems reasonable) and practice. This latter part is of great interest, unfortunately, however, detailed protocols (spermidine or no spermidine?) are not given, especially in the case of blunt end ligation. One has therefore to make one's own philosophy out of the rationale which is presented in the text. The present review could similarly go on with all general techniques detailed in this practical guide but let us stop here. In all it is a very useful guide, probably somewhat too dense, but certainly practical. It can be recommended for those who plan to use molecular genetics techniques. With the Introduction to Practical Molecular Biology, we are faced with a completely different type of practical guide: indeed rather than a quarto format like the preceding one it is a small octavo-sized one of about 100 pages of text. Its scope is mainly DNA and RNA preparation for hydridization studies. The content corresponds to a well presented laboratory notebook, i.e., with not much description of the concepts. As such it is only an introduction to the field for students starting molecular biology experiments, it should help students involved in courses, but probably not those who are working full time in a laboratory. In addition, many catalogues from profit-making laboratories usually contain similar free information. Thus it is not clear whether it will sell well. A. Danchin
Industrial Biocides, Vol. 23, by K.R. Payne. John Wiley & Sons, Chichester, 1988, I:P. 118, £ 37.50 The undesirable development of microorganisms in industrial ecosystems may be responsible for huge economic losses due to the degradation of product quality or to damage to materials. Uncontrolled presence of microorganisms can also pose health hazards in the hospital environment or in food processing plants. Commercial preparations including compounds called 'biocides" (probably several hundred molecules available) are currently used to control the development of bacteria and eukaryotic microorganisms. Industrial microbiologists are very often faced with the problem of obtaining information on the activity of such compounds or preparations because these data, if available, are widely scattered throughout the literature.
Book reviews A Practical Guide to Molecular Clo,fing, by B. Perbal. 2nd edn. John Wiley & Sons, Chichester, 1988, 812 pp., £ 32.50 Introduction to Practical Molecular Biology, by P.D. Darbre. John Wiley & Sons, Chichester, 1988, 117 pp., £ 9.95 Everybody aware of practical molecular genetics knows the blue cover of the 'Maniatis', and it is not a rare event that it has been stolen from a shelf... This may explain why this practical textbook has had many clones as did the IBM PC microcomputers. A Practical Guide to Molecular Cloning by B. Perbai (the second edition, already!) is such a clone, but different enough from its reference to be worth buying in addition to the holy book. Safety recommendations are present at the start of this big guide (chapter 2) and this is certainly a good starting point. Then follow all standard techniques of molecular biology: gel electrophoresis and filtration, ultracentrifugation, ion-exchange chromatography, autoradiography, DNA precipitation. Not much is said about/3-galactosidase or about affinity chromatography (for instance using this enzyme as a carrier protein); it is our hope that this hole will be filled in a future edition! Restriction and modifica6on are extensively developed, useful enzymes (and also others) are presented with their target sites, a lengthy table presents a usefnl nattprn nf huhrirl c~n,,~ne,~c rpcnltlna f r n r , ~ !igation generated after digestion with different restriction enzymes. Unfortunately, however, the understandable wish to be complete results in such a large table that it becomes practically useless. A table such as the one given in the well known Biolabs catalogue would be a significant improvement in that matter. At least isoschizomers should have been grouped (why separate BamHl and BstI?). But rather than go through each chapter (the book is over 800 pages long, including protocols), let's go through the protocols. The material is E. coli K12 or B, in general, with appropriate phage, cosmid or plasmid vectors. DNA preparation is therefore of major importance. A rapid procedure described for plasmid preparation involves, all standard ingredients (SDS, NaOH, and.., lysosyme, which could be omitted without any damage to the yield and purity), but adds several steps (dialysis on Millipore filters) which I have never personally used and might, perhaps, increase the digestion efficiency of some enzymes. A thorough analysis of restriction enzymes activity as a function of salt concentration will certainly be useful to many: it is always disappointing to obtain partial digestions on a sample containing only preciously little DNA. Another chapter (13) is devoted to an ever surprising step in molecular cloning: iigation. In this chapter we
are presented with theory (which I have not checked, but which seems reasonable) and practice. This latter part is of great interest, unfortunately, however, detailed protocols (spermidine or no spermidine?) are not given, especially in the case of blunt end ligation. One has therefore to make one's own philosophy out of the rationale which is presented in the text. The present review could similarly go on with all general techniques detailed in this practical guide but let us stop here. In all it is a very useful guide, probably somewhat too dense, but certainly practical. It can be recommended for those who plan to use molecular genetics techniques. With the Introduction to Practical Molecular Biology, we are faced with a completely different type of practical guide: indeed rather than a quarto format like the preceding one it is a small octavo-sized one of about 100 pages of text. Its scope is mainly DNA and RNA preparation for hydridization studies. The content corresponds to a well presented laboratory notebook, i.e., with not much description of the concepts. As such it is only an introduction to the field for students starting molecular biology experiments, it should help students involved in courses, but probably not those who are working full time in a laboratory. In addition, many catalogues from profit-making laboratories usually contain similar free information. Thus it is not clear whether it will sell well. A. Danchin
Industrial Biocides, Vol. 23, by K.R. Payne. John Wiley & Sons, Chichester, 1988, I:P. 118, £ 37.50 The undesirable development of microorganisms in industrial ecosystems may be responsible for huge economic losses due to the degradation of product quality or to damage to materials. Uncontrolled presence of microorganisms can also pose health hazards in the hospital environment or in food processing plants. Commercial preparations including compounds called 'biocides" (probably several hundred molecules available) are currently used to control the development of bacteria and eukaryotic microorganisms. Industrial microbiologists are very often faced with the problem of obtaining information on the activity of such compounds or preparations because these data, if available, are widely scattered throughout the literature.