Making PCR


Book Description

Making PCR is the fascinating, behind-the-scenes account of the invention of one of the most significant biotech discoveries in our time—the polymerase chain reaction. Transforming the practice and potential of molecular biology, PCR extends scientists' ability to identify and manipulate genetic materials and accurately reproduces millions of copies of a given segment in a short period of time. It makes abundant what was once scarce—the genetic material required for experimentation. Making PCR explores the culture of biotechnology as it emerged at Certus Corporation during the 1980s and focuses on its distinctive configuration of scientific, technical, social, economic, political, and legal elements, each of which had its own separate trajectory over the preceding decade. The book contains interviews with the remarkable cast of characters who made PCR, including Kary Mullin, the maverick who received the Nobel prize for "discovering" it, as well as the team of young scientists and the company's business leaders. This book shows how a contingently assembled practice emerged, composed of distinctive subjects, the site where they worked, and the object they invented. "Paul Rabinow paints a . . . picture of the process of discovery in Making PCR: A Story of Biotechnology [and] teases out every possible detail. . . . Makes for an intriguing read that raises many questions about our understanding of the twisting process of discovery itself."—David Bradley, New Scientist "Rabinow's book belongs to a burgeoning genre: ethnographic studies of what scientists actually do in the lab. . . . A bold move."—Daniel Zalewski, Lingua Franca "[Making PCR is] exotic territory, biomedical research, explored. . . . Rabinow describes a dance: the immigration and repatriation of scientists to and from the academic and business worlds."—Nancy Maull, New York Times Book Review




PCR Primer Design


Book Description

At the heart of most high-throughput methods is the technique of polymerase chain reaction (PCR). This book focuses on primer design, which is critical to both the efficiency and the accuracy of the PCR. With intricate descriptions of basic approaches as well as specialized methods, "PCR Primer Design" is an exceptional reference for all those involved in studying the genome.




PCR Protocols


Book Description

The correct procedures you need for frustration-free PCR methods and applications are contained in this complete, step-by-step, clearly written, inexpensive manual. - Avoid contamination--with specific instructions on setting up your lab - Avoid cumbersome molecular biological techniques - Discover new applications




Basic Science Methods for Clinical Researchers


Book Description

Basic Science Methods for Clinical Researchers addresses the specific challenges faced by clinicians without a conventional science background. The aim of the book is to introduce the reader to core experimental methods commonly used to answer questions in basic science research and to outline their relative strengths and limitations in generating conclusive data. This book will be a vital companion for clinicians undertaking laboratory-based science. It will support clinicians in the pursuit of their academic interests and in making an original contribution to their chosen field. In doing so, it will facilitate the development of tomorrow's clinician scientists and future leaders in discovery science. - Serves as a helpful guide for clinical researchers who lack a conventional science background - Organized around research themes pertaining to key biological molecules, from genes, to proteins, cells, and model organisms - Features protocols, techniques for troubleshooting common problems, and an explanation of the advantages and limitations of a technique in generating conclusive data - Appendices provide resources for practical research methodology, including legal frameworks for using stem cells and animals in the laboratory, ethical considerations, and good laboratory practice (GLP)




The Polymerase Chain Reaction


Book Description

James D. Watson When, in late March of 1953, Francis Crick and I came to write the first Nature paper describing the double helical structure of the DNA molecule, Francis had wanted to include a lengthy discussion of the genetic implications of a molecule whose struc ture we had divined from a minimum of experimental data and on theoretical argu ments based on physical principles. But I felt that this might be tempting fate, given that we had not yet seen the detailed evidence from King's College. Nevertheless, we reached a compromise and decided to include a sentence that pointed to the biological significance of the molecule's key feature-the complementary pairing of the bases. "It has not escaped our notice," Francis wrote, "that the specific pairing that we have postulated immediately suggests a possible copying mechanism for the genetic material." By May, when we were writing the second Nature paper, I was more confident that the proposed structure was at the very least substantially correct, so that this second paper contains a discussion of molecular self-duplication using templates or molds. We pointed out that, as a consequence of base pairing, a DNA molecule has two chains that are complementary to each other. Each chain could then act ". . . as a template for the formation on itself of a new companion chain, so that eventually we shall have two pairs of chains, where we only had one before" and, moreover, " ...




PCR Applications


Book Description

PCR is the most powerful technique currently used in molecular biology. It enables the scientist to quickly replicate DNA and RNA on the benchtop. From its discovery in the early 80's, PCR has blossomed into a method that enables everything from ready mutation of DNA/RNA to speedy analysis of tens of thousands of nucleotide sequences daily.PCR Applications examines the latest developments in this field. It is the third book in the series, building on the previous publications PCR Protocols and PCR Strategies. The manual discusses techniques that focus on gene discovery, genomics, and DNA array technology, which are contributing factors to the now-occurring bioinformatics boom.Key Features* Focuses on gene discovery, genomics, and DNA array technology* Covers quantitative PCR techniques, including the use of standards and kinetic analysisincludes statistical refinement of primer design parameters* Ilustrates techniques used in microscopic tissue samples, such as single cell PCR, whole cell PCR, laser capture microdissection, and in situ PCREntries provide information on:* Nomenclature* Expression* Sequence analysis* Structure and function* Electrophysiology* Parmacology* Information retrieval




PCR Methods in Foods


Book Description

This book will introduce non-molecular biologists to diagnostic PCR-based te- nologies for the detection of pathogens in foods. By the conclusion of this book, the reader should be able to: 1) understand the principles behind PCR including real-time; 2) know the basics involved in the design, optimization, and imp- mentation of PCR in food microbiology lab setting; 3) interpret results; 4) know limitations and strengths of PCR; and 5) understand the basic principles behind a new fledgling technology, microarrays and its potential applications in food microbiology. This book will provide readers with the latest information on PCR and microarray based tests and their application towards the detection of bacterial, protozoal and viral pathogens in foods. Figures, charts, and tables will be used, where appropriate, to help illustrate concepts or provide the reader with useful information or resources as an important starting point in bringing molecular diagnostics into the food microbiology lab. This book is not designed to be a “cookbook”PCR manual with recipes and step-by-step instructions but rather serve as a primer or resource book for students, faculty, and other professionals interested in molecular biology and its integration into food safety. v Table of Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Chapter 1. PCR Basics Amanda Fairchild, M. S. , Margie D. Lee DVM, Ph. D. , and John J. Maurer, Ph. D. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 Chapter 2. The Mythology of PCR: A Warning to the Wise John J. Maurer, Ph. D. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 Chapter 3.




A Machine to Make a Future


Book Description

A Machine to Make a Future represents a remarkably original look at the present and possible future of biotechnology research in the wake of the mapping of the human genome. The central tenet of Celera Diagnostics--the California biotech company whose formative work during 2003 is the focus of the book--is that the emergent knowledge about the genome, with its profound implications for human health, can now be turned into a powerful diagnostic apparatus--one that will yield breakthrough diagnostic and therapeutic products (and, potentially, profit). Celera's efforts--assuming they succeed--may fundamentally reshape the fabric of how health and health care are understood, practiced, and managed. Presenting a series of interviews with all of the key players in Celera Diagnostics, Paul Rabinow and Talia Dan-Cohen open a fascinating window on the complexity of corporate scientific innovation. This marks a radical departure from other books on the biotech industry by chronicling the vicissitudes of a project during a finite time period, in the words of the actors themselves. Ultimately, the authors conclude, Celera Diagnostics is engaged in a future characterized not by geniuses and their celebrated discoveries but by a largely anonymous and widely distributed profusion of data and results--a "machine to make a future." In their new afterword, Rabinow and Dan-Cohen revisit Celera Diagnostics as its mighty machine grinds along, wondering, along with the scientists, "what constitutes success and what constitutes failure?" The pathos of the situation turns on how one poses the question as much as how one answers it.




PCR Cloning Protocols


Book Description

PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination, and to clone the challenging uncharacterized DNA flanking a known DNA fragment.




Principles and Technical Aspects of PCR Amplification


Book Description

Kary Mullis was awarded a Nobel Prize for inventing the PCR technique more than a decade ago in 1993. Since its "discovery", multiple adaptations and variations of the standard PCR technique have been described. This publication aims to provide the reader with a guide to the standard PCR technique and its many available variants, with particular emphasis being placed on the role of these PCR techniques in the clinical diagnostic laboratory (the central theme of this book).