Can't clone, won't clone

2001 ◽  
Vol 114 (10) ◽  
pp. 1797-1798
Author(s):  
K. Plant

Essential Molecular Biology, Vol. 1, 2nd edn edited by T. A. Brown Oxford University Press (2000) 240 pages. ISBN 0–19-963642-7 pound29.95 I have heard it said (though not to my face) that practical molecular biology is somewhat akin to cookery, and I have to admit (though not to my fellow Molecular Biologists) that there is an element of truth in this. Of course, our ovens are cooler, and our pie dishes smaller, but often it is a case of mixing ingredients in the right proportions and baking at 37 degrees C for an hour. In this book Brown becomes the Delia Smith of molecular biology, starting with how to boil an egg, before proceeding to more complex recipes. It is utterly and unashamedly aimed at the complete novice. As more and more branches of biology use molecular techniques, and as a constant flow of graduates take up the yoke of research, there will always be a demand for this type of manual. Of course, it is possible to find variants of a lot of these methods on the Internet, but, as these often include only the protocol, the complete novice is probably better off with a specifically designed manual. Another option is to go for the kit approach, but, in the same way as opening a tin of beans doesn't make you a chef, I whole heartedly agree with Brown when he says, ‘do not get the idea that using kits is the same as being a molecular biologist.’ In most branches of biology a bit of genuine molecular expertise can only enhance one's future job prospects! One of the things I liked about this book is its no-nonsense style, particularly those chapters written by the Editor. There is plenty of sound advice, not just on the molecular techniques but on how to be a good scientist in general. Although the advice starts with the basics, it isn't patronising to those experienced in other fields. One piece of advice that particularly tickled me was that, if your hand is too unsteady to load a gel, you should give up caffeine; I'm not sure whether the pain would be worth the gain! The first chapter deals with all the basic issues, from planning (not just how to do it, but is it worthy of your time, which is something we should all think about occasionally) to safety (which nasties you'll be using, what precautions to take, with internet sites referenced to fill in the details) and what equipment you'll need to run the experiments. In a nutshell, the rest of the book deals with microbiology for molecular biologists and molecular biology for everyone else. This includes DNA and RNA isolation, electrophoresis and cloning (generating, propagating and identifying recombinant DNA molecules, not the Dolly-the-sheep variety). There is a second volume to the set, which (based on the contents of the first edition) should cover making and screening libraries, the polymerase chain reaction, sequencing and gene expression studies. Bear in mind that to get very far you will need to buy the second volume, which is not yet published. It has been more than a decade since the first edition of this well-known and respected manual was published; so one would think its first update is about due. However, compared with the first edition, most chapters have very few changes. This is probably in the nature of such a basic manual - for example, good microbial practice doesn't change much. Only a couple of chapters have been extensively rewritten; those describing DNA extraction now include more recent resin-based methods. So to the crux of the matter: would I recommend buying it? Well, if you're a complete novice with little backup, I definitely think it is worth investing in a decent manual, and this one does have a nice comfortable feel to it. If you've already got a copy of the previous edition and are wondering whether to upgrade, I would say that the few improvements in these very basic techniques do not really make it worth spending the pound30 that this volume costs. That said, I rather suspect that the second volume, which deals with more complex techniques, will show far more technical advances and should complete your progression from culinary incompetence to cordon bleu.

2000 ◽  
Vol 113 (22) ◽  
pp. 3887-3888
Author(s):  
K.E. Plant

Basic Techniques in Molecular Biology by Stefan Surzycki Springer-Verlag (2000) pp. 434. ISBN 3–540-66678-8 pound44.50/$79.95 This laboratory manual differs from many of the numerous others currently on the market in that it explains the ‘Whys’ as well as the ‘Hows’ of the most commonly used molecular techniques. The book describes DNA and RNA isolation, electrophoresis, blotting and hybridisation, cloning, sequencing and of course PCR. Each chapter consists of a description of the principles involved, a schematic outline of the procedure, 3 or 4 detailed protocols and a trouble-shooting guide. The book is principally aimed at students and others who are relatively new to practical molecular biology and the emphasis is very definitely on making techniques accessible to these people. I rather enjoyed reviewing Dr Surzycki's manual. It is absolutely jam packed full of those little titbits of information that friendly Post Docs occasionally feel inclined to share with students. If you love to delve into the details of what you are doing, you want to know why you're using ammonium and not sodium acetate or can't decide between a phenol or a chloroform extraction, then I think you'll enjoy perusing this book too. I defy anybody to read this manual without picking up some little snippet of information they didn't previously know. And even if you did know it, it's nice to have it before you in black and white instead of as a vague memory of something somebody once told you! As the author admits, there is a degree of repetition throughout the book. What this means is that most of the chapters essentially stand alone, so there is no hunting around for recipes, but of course it also pushes up the size (and therefore the price) of the book. Each chapter also has a trouble shooting guide complete with ‘Recovery Protocols’, which could be very handy for the blundering novice. For the less brave there are also lists of suppliers of kits for many of the procedures, complete with the company's website, a brief description of the principles the kit relies on and the odd favourable opinion. The author tries to give users enough background information to allow them to modify and design new protocols, and he certainly succeeds in that respect. Lessons learned in one technique can always be applied to another, and if you can understand exactly what a particular procedure does then trouble shooting those other techniques suddenly becomes a lot easier. So as you'll have gathered by now I liked this book, and really the only criticism I can level at it is that it is rather restricted in the range of techniques it covers. For anyone who is at all serious about molecular biology it is only ever going to serve as a starting place and there is a wealth of other techniques out there which this book doesn't even touch upon. That said there are also plenty of more specialised manuals out there too, and this particular book more than adequately fills the niche it is intended to cover.


2005 ◽  
Vol 347 (1) ◽  
pp. 156-158 ◽  
Author(s):  
Olha Khymenets ◽  
Jordi Ortuño ◽  
Montserrat Fitó ◽  
Ma Isabel Covas ◽  
Magí Farré ◽  
...  

2005 ◽  
Vol 17 (9) ◽  
pp. 105
Author(s):  
J. Liew ◽  
A. E. Drummond ◽  
M. E. Jones ◽  
M. Poutanen ◽  
J. K. Findlay

Aromatase, the product of the Cyp 19 gene, converts androgens to estrogens. The role of estrogens within the ovary has recently been revisited; using the aromatase knockout (ArKO) mouse, we investigated the effect of estrogen deficiency on ovarian function. We now have an aromatase overexpressing (AROM+) female mouse model with elevated levels of estrogens. These mice were fertile and bred with FVB/N wildtype (WT) males, the AROM+ male being infertile. In this study we characterised the reproductive phenotype of the female AROM+ mouse. 5 WT and 10 AROM+ mice, 22–27 weeks of age were used in the study. The mice were subject to vaginal smears and killed during estrus. The ovaries, uterine horns and gonadal fat were collected and weighed. One ovary and the uterine horns were fixed in formalin for histological assessment, while the other ovary was snap frozen in Ultraspec solution for RNA isolation and gene expression studies. Serum was collected for hormone measurements. All AROM+ mice exhibited an abnormal pattern of cycling that in general, alternated between estrus and post-estrus. AROM+ mice were significantly heavier than their WT counterparts (WT 35.28 ± 2.89 g v. AROM+ 43.38 ± 2.11 g, P < 0.05). Ovarian, uterine and gonadal fat pad weights were not significantly different between the 2 groups (ovary: WT 17.4 ± 1.14 mg v. AROM+ 17.9 ± 0.06 mg; uterine horns: WT 89.7 ± 11.40 mg v. AROM+ 92.1 ± 6.64 mg; gonadal fat pads: WT 2.47 ± 0.62 g v. AROM+ 3.46±0.26 g). Histological, gene expression and hormone analyses are in progress. Our preliminary analyses indicated no significant effect of excess estrogen on ovarian, uterine and gonadal fat pad weights, despite the AROM+ mice being heavier. It remains to be determined as to whether the ovaries and uterine horns are histologically normal. Supported by the NHMRC (Regkeys 241000, 338510, 198705)


2004 ◽  
Vol 3 (1) ◽  
pp. 31-48 ◽  
Author(s):  
Alison E. Gammie ◽  
Naz Erdeniz

This work describes the project for an advanced undergraduate laboratory course in cell and molecular biology. One objective of the course is to teach students a variety of cellular and molecular techniques while conducting original research. A second objective is to provide instruction in science writing and data presentation by requiring comprehensive laboratory reports modeled on the primary literature. The project for the course focuses on a gene, MSH2, implicated in the most common form of inherited colorectal cancer. Msh2 is important for maintaining the fidelity of genetic material where it functions as an important component of the DNA mismatch repair machinery. The goal of the project has two parts. The first part is to create mapped missense mutation listed in the human databases in the cognate yeast MSH2 gene and to assay for defects in DNA mismatch repair. The second part of the course is directed towards understanding in what way are the variant proteins defective for mismatch repair. Protein levels are analyzed to determine if the missense alleles display decreased expression. Furthermore, the students establish whether the Msh2p variants are properly localized to the nucleus using indirect immunofluorescence and whether the altered proteins have lost their ability to interact with other subunits of the MMR complex by creating recombinant DNA molecules and employing the yeast 2-hybrid assay.


2017 ◽  
Vol 42 (4) ◽  
Author(s):  
Synda Chenenaoui ◽  
Samia Daldoul ◽  
Ahmed Mliki

AbstractObjectives:Grapevine root system plays a great role in sensing and adapting to abiotic and biotic stresses. Identification of candidate genes involved in the tolerance to abiotic stress is becoming a crucial strategy to select and breed resilient genotypes. However, obtaining high quality RNA from grapevine roots under hydroponic culture is difficult. Hence, we have developed a new extraction procedure to improve RNA quality for root gene expression studies.Methods:Conventional RNA extraction methods using CTAB are not suitable for gene expression studies and need to be improved. Here we report the application of a CTAB- based method for RNA extraction using an additional clean-up purification step.Results:The RIN value of the resulting RNA indicated that our procedure allowed the purification of high RNA quality and quantity. Hence, the clean-up purification step efficiently eliminated contaminants which inhibit downstream applications. Derived RNA was successfully used for differential gene expression analysis in salt stressed grapevine by Northern Blot hybridizations.Conclusion:In this study, we developed an efficient RNA isolation protocol from hydroponic cultivated grapevine roots which yielded RNA suitable for gene expression studies. This will open large perspectives in grapevine functional genomics with the identification of pertinent genes of agronomic interest.


CNS Spectrums ◽  
1997 ◽  
Vol 2 (9) ◽  
pp. 35-41
Author(s):  
Silvia Marracci ◽  
Donatella Marazziti ◽  
Michela Ori ◽  
Irma Nardi

AbstractMolecular biology techniques are used widely in the study of central nervous system structures and functions. The possibilities offered by such methodologies are attracting an increasing number of researchers for their versatility and for their promise to permit the investigation of different proteins, step by step, from gene transcription to posttranscriptional processes. Of particular interest is the study of the enzymes responsible for the synthesis and catabolism of neurotransmitters, as well as neuroreceptors.Today, a general knowledge of such methods and of how they can be used in CNS studies is fundamental for psychiatrists, even if not directly involved in basic research, since it can be easily predicted that the next century will be characterized by an increasing application of these approaches to biological psychiatry. In this article, we describe gene expression methods and the aim of gene expression studies. We also present some general information on the various steps and techniques used for different purposes in biological psychiatry.


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