This first book on the market covers the many new and important RNA species discovered over the past five years, explaining current methods for the enrichment, separation and purification of these novel RNAs. Building up from general principles of RNA biochemistry and biophysics, this book addresses the practical aspects relevant to the laboratory researcher throughout, while discussing the performance and potential problems of the methods discussed. An appendix contains a glossary with the important terms and techniques used in RNA analysis. By explaining the basic and working principles of the methods, the book allows biochemists and molecular biologists to gain much more expertise than by simply repeating a pre-formulated protocol, enabling them to select the procedure and materials best suited to the RNA analysis task at hand. As a result, they will be able to develop new protocols where needed and optimize and fine-tune the general purpose standard protocols that come with the purification equipment and instrumentation.

Ribonucleic acids are central to cellular and molecular processes and perform vital functions in both structural and functional roles. RNA molecules form the bridge between the stable genetic information contained within DNA and enzymes and proteins that carry out much of the metabolism within the cell. Many of the sites of protein synthesis, the ribosomes within the cell, are composed of these ribonucleic acids as are the tRNA molecules that deliver the amino acid building blocks to the ribosomes. Of all the RNA species, the nucleic acid intermediate, messenger RNA, is a desirable source of material to biologists, since this reflects much of, what ultimately, is translated into enzymes and proteins. In order to determine the qualitative and quantitative changes in mRNA expression, a vast number of molecular biological techniques have been developed. Key molecular methods that provide the means to initially isolate and analyze RNA molecules are the focus of this volume. In putting together this collection of protocols, we have tried to provide techniques that are most applicable and widely used. In particular, there are a number of iso- tion techniques included that have been developed, modified, or adapted to enable extraction from a variety of cell types, organisms, or subcellular organelles. Successful isolation of intact RNA is an essential starting point for any sub- quent analysis. This is why we have aimed to make this section comprehensive. The analysis of RNA is the focus of the following chapters.

This book examines a wide range of techniques on RNA extraction, detection, quantification, visualization, and genome-wide profiling, from conventional methods to state-of-the-art high throughput approaches.

The second edition of a highly acclaimed handbook and ready reference. Unmatched in its breadth and quality, around 100 specialists from all over the world share their up-to-date expertise and experiences, including hundreds of protocols, complete with explanations, and hitherto unpublished troubleshooting hints. They cover all modern techniques for the handling, analysis and modification of RNAs and their complexes with proteins. Throughout, they bear the practising bench scientist in mind, providing quick and reliable access to a plethora of solutions for practical questions of RNA research, ranging from simple to highly complex. This broad scope allows the treatment of specialized methods side by side with basic biochemical techniques, making the book a real treasure trove for every researcher experimenting with RNA.

This book is designed to be a practical progression of experimental techniques an investigator may follow when embarking on a biochemical project. The protocols may be performed in the order laid out or may be used inde pendently. The aim of the book is to assist a wide range of researchers. from the novice to the frustrated veteran, in the choice and design of experiments that are to be performed to provide answers to specific questions. The manual describes standard techniques that have been shown to work, as well as some newer ones that are beginning to prove important. By following the promi nently numbered steps. you can work your way through any protocol. whether it's a new technique or a task you've done before for which you need a quick review or updated methodology. This manual will assist the experimentalist in designing properly controlled experiments. There will be no advice for dealing with specific pieces of equip ment other than encouragement to read the manual, if you can find it. Through out all manipulations try to be objective. Be on the lookout for unexpected findings. You will learn the most from unexpected results. and they are often the beginning of the next project. It is never possible to record too much in your lab notebook. Do not get discouraged. Remember, things will not always run smoothly.

An Indispensable Roadmap for Nucleic Acid Preparation Although Friedrich Miescher described the first isolation of nucleic acid in 1869, it was not until 1953 that James Watson and Francis Crick successfully deciphered the structural basis of DNA duplex. Needless to say, in the years since, enormous advances have been made in the study of nucleic a

This laboratory guide represents a growing collection of tried, tested and optimized laboratory protocols for the isolation and characterization of eukaryotic RNA, with lesser emphasis on the characterization of prokaryotic transcripts. Collectively the chapters work together to embellish the RNA story, each presenting clear take-home lessons, liberally incorporating flow charts, tables and graphs to facilitate learning and assist in the planning and implementation phases of a project.RNA Methodologies, 3rd edition includes approximately 30% new material, including chapters on the more recent technologies of RNA interference including: RNAi; Microarrays; Bioinformatics. It also includes new sections on: new and improved RT-PCR techniques; innovative 5' and 3' RACE techniques; subtractive PCR methods; methods for improving cDNA synthesis.* Author is a well-recognized expert in the field of RNA experimentation and founded Exon-Intron, a well-known biotechnology educational workshop center * Includes classic and contemporary techniques * Incorporates flow charts, tables, and graphs to facilitate learning and assist in the planning phases of projects

A comprehensive treasury of all the key molecular biology methods-ranging from DNA extraction to gene localization in situ-needed to function effectively in the modern laboratory. Each of the 120 highly successful techniques follows the format of the much acclaimed Methods in Molecular BiologyOao series, providing an introduction to the scientific basis of each technique, a complete listing of all the necessary materials and reagents, and clear step-by-step instruction to permit error-free execution. Included for each technique are notes about pitfalls to avoid, troubleshooting tips, alternate methods, and explanations of the reasons for certain steps-all key elements contributing significantly to success or failure in the lab. The Nucleic Acid Protocols Handbook constitutes today's most comprehensive collection of all the key classic and cutting-edge techniques for the successful isolation, analysis, and manipulation of nucleic acids by both experienced researchers and those new to the field."

The importance of accurate sample preparation techniques cannot be overstated--meticulous sample preparation is essential. Often overlooked, it is the midway point where the analytes from the sample matrix are transformed so they are suitable for analysis. Even the best analytical techniques cannot rectify problems generated by sloppy sample pretreatment. Devoted entirely to teaching and reinforcing these necessary pretreatment steps, Sample Preparation Techniques in Analytical Chemistry addresses diverse aspects of this important measurement step. These include: * State-of-the-art extraction techniques for organic and inorganic analytes * Sample preparation in biological measurements * Sample pretreatment in microscopy * Surface enhancement as a sample preparation tool in Raman and IR spectroscopy * Sample concentration and clean-up methods * Quality control steps Designed to serve as a text in an undergraduate or graduate level curriculum, Sample Preparation Techniques in Analytical Chemistry also provides an invaluable reference tool for analytical chemists in the chemical, biological, pharmaceutical, environmental, and materials sciences.