Tricks of the trade: a beginner's guide to practical molecular biology

2000 ◽  
Vol 113 (22) ◽  
pp. 3887-3888
Author(s):  
K.E. Plant
Keyword(s):  
Molecular Biology ◽  
Rna Isolation ◽  
Lessons Learned ◽  
Molecular Techniques ◽  
Background Information ◽  
Laboratory Manual ◽  
Black And White ◽  
Trouble Shooting ◽  
Dna And Rna ◽  
Recovery Protocols

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.

Can't clone, won't clone

2001 ◽  
Vol 114 (10) ◽  
pp. 1797-1798
Author(s):  
K. Plant
Keyword(s):  
Molecular Biology ◽  
Recombinant Dna ◽  
Rna Isolation ◽  
Molecular Techniques ◽  
Dna And Rna ◽  
Expression Studies ◽  
Previous Edition ◽  
Technical Advances ◽  
Gene Expression Studies ◽  
The Right

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.

scholarly journals Role of Laboratory Medicine in SARS-CoV-2 Diagnostics. Lessons Learned from a Pandemic

Healthcare ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 915
Author(s):  
Irena Duś-Ilnicka ◽  
Aleksander Szymczak ◽  
Małgorzata Małodobra-Mazur ◽  
Miron Tokarski
Keyword(s):  
Molecular Biology ◽  
Real Time ◽  
Real Time Pcr ◽  
Laboratory Medicine ◽  
Lessons Learned ◽  
Diagnostic Methods ◽  
Medical Community ◽  
Medical Diagnostic ◽  
Novel Coronavirus ◽  
Available Information

Since the 2019 novel coronavirus outbreak began in Wuhan, China, diagnostic methods in the field of molecular biology have been developing faster than ever under the vigilant eye of world’s research community. Unfortunately, the medical community was not prepared for testing such large volumes or ranges of biological materials, whether blood samples for antibody immunological testing, or salivary/swab samples for real-time PCR. For this reason, many medical diagnostic laboratories have made the switch to working in the field of molecular biology, and research undertaken to speed up the flow of samples through laboratory. The aim of this narrative review is to evaluate the current literature on laboratory techniques for the diagnosis of SARS-CoV-2 infection available on pubmed.gov, Google Scholar, and according to the writers’ knowledge and experience of the laboratory medicine. It assesses the available information in the field of molecular biology by comparing real-time PCR, LAMP technique, RNA sequencing, and immunological diagnostics, and examines the newest techniques along with their limitations for use in SARS-CoV-2 diagnostics.

scholarly journals Modernizing the Toolkit for Arthropod Bloodmeal Identification

Insects ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 37
Author(s):  
Erin M. Borland ◽  
Rebekah C. Kading
Keyword(s):  
Nuclear Dna ◽  
Point Of Care ◽  
Isotope Analysis ◽  
Digital Pcr ◽  
Lessons Learned ◽  
Molecular Techniques ◽  
Melting Analysis ◽  
Field Deployment ◽  
Blood Meal Identification ◽  
Bloodmeal Identification

Understanding vertebrate–vector interactions is vitally important for understanding the transmission dynamics of arthropod-vectored pathogens and depends on the ability to accurately identify the vertebrate source of blood-engorged arthropods in field collections using molecular methods. A decade ago, molecular techniques being applied to arthropod blood meal identification were thoroughly reviewed, but there have been significant advancements in the techniques and technologies available since that time. This review highlights the available diagnostic markers in mitochondrial and nuclear DNA and discusses their benefits and shortcomings for use in molecular identification assays. Advances in real-time PCR, high resolution melting analysis, digital PCR, next generation sequencing, microsphere assays, mass spectrometry, and stable isotope analysis each offer novel approaches and advantages to bloodmeal analysis that have gained traction in the field. New, field-forward technologies and platforms have also come into use that offer promising solutions for point-of-care and remote field deployment for rapid bloodmeal source identification. Some of the lessons learned over the last decade, particularly in the fields of DNA barcoding and sequence analysis, are discussed. Though many advancements have been made, technical challenges remain concerning the prevention of sample degradation both by the arthropod before the sample has been obtained and during storage. This review provides a roadmap and guide for those considering modern techniques for arthropod bloodmeal identification and reviews how advances in molecular technology over the past decade have been applied in this unique biomedical context.

scholarly journals Visual pH Sensors: From a Chemical Perspective to New Bioengineered Materials

Molecules ◽  
2021 ◽  
Vol 26 (10) ◽  
pp. 2952
Author(s):  
Luigi Di Costanzo ◽  
Barbara Panunzi
Keyword(s):  
Ph Monitoring ◽  
Three Dimensional ◽  
Lessons Learned ◽  
Background Information ◽  
Biological Macromolecules ◽  
Ph Sensing ◽  
Ph Sensors ◽  
Cellular Functions ◽  
Metal Organic

Many human activities and cellular functions depend upon precise pH values, and pH monitoring is considered a fundamental task. Colorimetric and fluorescence sensors for pH measurements are chemical and biochemical tools able to sense protons and produce a visible signal. These pH sensors are gaining widespread attention as non-destructive tools, visible to the human eye, that are capable of a real-time and in-situ response. Optical “visual” sensors are expanding researchers’ interests in many chemical contexts and are routinely used for biological, environmental, and medical applications. In this review we provide an overview of trending colorimetric, fluorescent, or dual-mode responsive visual pH sensors. These sensors include molecular synthetic organic sensors, metal organic frameworks (MOF), engineered sensing nanomaterials, and bioengineered sensors. We review different typological chemical entities of visual pH sensors, three-dimensional structures, and signaling mechanisms for pH sensing and applications; developed in the past five years. The progression of this review from simple organic molecules to biological macromolecules seeks to benefit beginners and scientists embarking on a project of pH sensing development, who needs background information and a quick update on advances in the field. Lessons learned from these tools will aid pH determination projects and provide new ways of thinking for cell bioimaging or other cutting-edge in vivo applications.

scholarly journals High-quality total RNA isolation from melon (Cucumis melo L.) fruits rich in polysaccharides

2017 ◽  
Vol 38 (4) ◽  
pp. 2201 ◽  
Author(s):  
Gabrielle Silveira de Campos ◽  
Ricardo Antônio Ayub ◽  
Rafael Mazer Etto ◽  
Carolina Weigert Galvão ◽  
Marília Aparecida Stroka ◽  
...  
Keyword(s):  
Rna Extraction ◽  
Sodium Perchlorate ◽  
Rna Isolation ◽  
World Market ◽  
Molecular Techniques ◽  
Guanidine Thiocyanate ◽  
High Quality ◽  
Total Rna ◽  
High Concentrations ◽  
Cucumis Melo L

Melon, a member of the family Cucurbitaceae, is the fourth most important fruit in the world market and, on a volume basis, is Brazil’s main fresh fruit export. Many molecular techniques used to understand the maturation of these fruits require high concentrations of highly purified RNA. However, melons are rich in polyphenolic compounds and polysaccharides, which interfere with RNA extraction. This study aimed to determine the most appropriate method for total RNA extraction from melon fruits. Six extraction buffers were tested: T1) guanidine thiocyanate/phenol/chloroform; T2) sodium azide/?-mercaptoethanol; T3) phenol/guanidine thiocyanate; T4) CTAB/PVP/?-mercaptoethanol; T5) SDS/sodium perchlorate/PVP/?-mercaptoethanol, and T6) sarkosyl/PVP/guanidine thiocyanate, using the AxyPrepTM Multisource Total RNA Miniprep Kit. The best method for extracting RNA from both mature and green fruit was based on the SDS/PVP/?-mercaptoethanol buffer, because it rapidly generated a high quality and quantity of material. In general, higher amounts of RNA were obtained from green than mature fruits, probably due to the lower concentration of polysaccharides and water. The purified material can be used as a template in molecular techniques, such as microarrays, RT-PCR, and in the construction of cDNA and RNA-seq data.

Mitochondrial DNA and RNA isolation from small amounts of potato tissue

2001 ◽  
Vol 19 (1) ◽  
pp. 67-67 ◽  
Author(s):  
Nunzia Scotti ◽  
Teodoro Cardi ◽  
Laurence Marechaldrouard
Keyword(s):  
Mitochondrial Dna ◽  
Rna Isolation ◽  
Potato Tissue ◽  
Dna And Rna

From Victim to Witness and the Challenges of Sexual Violence Testimony

2020 ◽  
pp. 118-124
Author(s):  
Jaqueline Moudeïna
Keyword(s):  
Sexual Violence ◽  
International Crime ◽  
Lessons Learned ◽  
Background Information ◽  
Amicus Curiae

This chapter discusses how Jacqueline Moudeïna, the first woman to practice law in Chad, became connected to Hissène Habré's victims, as well as how she organized and prepared them for trial. Moudeïna coordinated the team of lawyers who represented nearly 4,000 victims in the trial of the former Chadian President Hissène Habré before the Extraordinary African Chambers (EAC). The preparation of the victims was a long term undertaking and a day-to-day effort. It was very difficult for the victims to testify, but the majority stood their ground. The chapter then looks at how Moudeïna convinced the victims of sexual violence to testify. It also considers the lessons learned with respect to enabling disclosure of sexual testimony, which became central to the trial. It should be noted that the amicus curiae brief on crimes of sexual violence was of great importance as background information. It helped clarify rape as an international crime in itself.

scholarly journals André Boivin: A pioneer in endotoxin research and an amazing visionary during the birth of molecular biology

2019 ◽  
Vol 26 (3) ◽  
pp. 165-171
Author(s):  
Jean-Marc Cavaillon
Keyword(s):  
Molecular Biology ◽  
Premature Death ◽  
Biological Chemistry ◽  
Scientific Contribution ◽  
Bacterial Dna ◽  
Dna And Rna ◽  
Assistant Director ◽  
Pyrimidine Bases ◽  
Carbon Molecules ◽  
Set Up

André Boivin (1895–1949) started his career in Marseille as a biochemist. Soon after the discovery of insulin, he worked on its purification, allowing for the treatment of local patients. He later moved to Strasbourg and set-up a microtitration technique of small carbon molecules and a method for quantifying purine and pyrimidine bases. His main scientific contribution occurred in Bucharest, where he was recruited to organize the teaching of medicinal chemistry. Together with Ion and Lydia Mesrobeanu, at the Cantacuzene Institute, they were the first to characterize the biochemical nature of endotoxins, which he termed the “glucido-lipidic antigen.” After joining the Institut Pasteur annex near Paris, he worked with Gaston Ramon pursuing his research on smooth and rough LPS. Additionally, with Albert Delaunay, he researched the formation of exotoxins and antibodies (Abs). He was nominated assistant-director of the Institut Pasteur in 1940. He initiated research on bacterial DNA and RNA, and was the first to hypothesize on how RNA fits into gene function. In 1947 he moved for a second time to Strasbourg, accepting a position as a Professor of Biological Chemistry. After his premature death at the age of 54, the French academies mourned his loss and recognized him as one of their outstanding masters of biochemistry, microbiology, immunology, and molecular biology.

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