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PCR Strategies expands and updates the landmark volume PCR Protocols. It is a companion laboratory manual that provides a completely new set of up-to-date strategies and protocols for getting the most from PCR. The editors have organized the book into four sections, focusing on principles, analyses, research applications, and alternative strategies for a wide variety of basic and clinical needs. If you own PCR Protocols, you will want PCR Strategies. If you don't own PCR Protocols, you will want to buy both! - Concepts explained - Methods detailed - Trouble-shooting emphasized - Novel applications highlighted - Key concepts for PCR - Analysis of PCR products - Research applications - Alternative amplification strategies
This second volume focuses on PCR methods and PCR application specificities to the biotechnology and bioengineering field. New and updated chapters detail real-time PCR protocols, synthetic biology applications, pathogen detection, microfluidics, digital, multiplex detection recent advances. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, PCR: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
With a variety of detection chemistries, an increasing number of platforms, multiple choices for analytical methods and the jargon emerging along with these developments, real-time PCR is facing the risk of becoming an intimidating method, especially for beginners. Real-time PCR provides the basics, explains how they are exploited to run a real-time PCR assay, how the assays are run and where these assays are informative in real life. It addresses the most practical aspects of the techniques with the emphasis on 'how to do it in the laboratory'. Keeping with the spirit of the Advanced Methods Series, most chapters provide an experimental protocol as an example of a specific assay.
The volumes in this series include contemporary techniques significant to a particular branch of neuroscience. They are an invaluable aid to the student as well as the experienced researcher not only in developing protocols in neuroscience but in disciplines where research is becoming closely related to neuroscience. Each volume of Methods in Neurosciences contains an index, and each chapter includes references. Dr. Conn became Editor-in-Chief of the series beginning with Volume 15, so each subsequent volume could be guest-edited by an expert in that specific field. This further strengthens the depth of coverage in Methods in Neurosciences for students and researchers alike. - Direct application of PCR to fresh or frozen clinical specimens (e.g., blood and solid tissue) - Complete retrieval of novel expressed genes by PCR without screening a library - Quantitation by PCR - Mutagenesis by PCR - PCR in AIDS research - Simple and effective protocols for PCR on archival specimens
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
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.
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
Geneticists and molecular biologists have been interested in quantifying genes and their products for many years and for various reasons (Bishop, 1974). Early molecular methods were based on molecular hybridization, and were devised shortly after Marmur and Doty (1961) first showed that denaturation of the double helix could be reversed - that the process of molecular reassociation was exquisitely sequence dependent. Gillespie and Spiegelman (1965) developed a way of using the method to titrate the number of copies of a probe within a target sequence in which the target sequence was fixed to a membrane support prior to hybridization with the probe - typically a RNA. Thus, this was a precursor to many of the methods still in use, and indeed under development, today. Early examples of the application of these methods included the measurement of the copy numbers in gene families such as the ribosomal genes and the immunoglo bulin family. Amplification of genes in tumors and in response to drug treatment was discovered by this method. In the same period, methods were invented for estimating gene num bers based on the kinetics of the reassociation process - the so-called Cot analysis. This method, which exploits the dependence of the rate of reassociation on the concentration of the two strands, revealed the presence of repeated sequences in the DNA of higher eukaryotes (Britten and Kohne, 1968). An adaptation to RNA, Rot analysis (Melli and Bishop, 1969), was used to measure the abundance of RNAs in a mixed population.
This volume provides an overview on design PCR primers for successful DNA amplification. Chapters focus on primer design strategies for quantitative PCR, in silico PCR primer design, and primer design using software. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, PCR Primer Design, Second Edition seeks to aid molecular biology students, researchers, professors and PCR enthusiasts.
The analysis of DNA sequence polymorphisms and mutations is of central importance in understanding biological systems. This book is devoted to the experimental analysis of DNA and presents easy-to-follow protocols. Various techniques from the simple to the highly complex are detailed in this volume, providing a wide spectrum of available methods and practical advice. The methods are described in terms of: History and background Principles and theory Equipment and reagents Protocols Troubleshooting Applications Improvements Results Comparisons with other methods Future prospects and developments This is an essential manual for researchers working in human, animal, or plant molecular genetics and is particularly valuable for hospital and commercial laboratories.