This is not just a cookbook for real-time quantitative PCR (qPCR). Admittedly, there are lots of recipes from distinguished contributors and Bustin has attempted to collect, sift through and rationalize the vast amount of information that is available on this subject. And yes, this book was conceived as a comprehensive hands-on manual to allow both the novice researcher and the expert to set up and carry out qPCR assays from scratch.
However, this book also sets out to explain as many features of qPCR as possible, provide alternative viewpoints and methods and, perhaps most importantly, aims to stimulate the researcher into generating, interpreting and publishing data that are reproducible, reliable and biologically meaningful.
In this book, leading scientists from academia as well as biotech and pharma companies introduce the revolutionary concept of designing RNA and DNA oligonucleotides with novel functions by in vitro selection. These functions comprise high affinity binding (aptamers), catalytic activity (ribozymes and deoxyribozymes) or combinations of binding and catalytic properties (aptazymes).
Basic concepts and technologies describing in detail how these functional oligonucleotides can be identified are presented. Numerous examples demonstrate the versatility of in vitro selected oligonucleotides. Special emphasis has been put on a section that shows the broad applicability of aptamers, e. g. in target validation, for analytics, or as new therapeutics. This first overview in the field is of prime interest for a broad audience of scientists both in academia and in industry who wish to expand their knowledge on the potential of new oligonucleotide functions and their applications.
This resource presents protocols for isolating a gene, cloning and characterizing it, expressing its encoded protein, and purifying and characterizing the protein′s physical properties. It includes background and procedures and is structured around 20 experiments that demonstrate how to prepare, manipulate, and analyze plasmids, produce fusion proteins in bacteria, and purify these proteins based on chemical properties or substrate affinities.
It describes topics such as the use of antibodies and techniques developed to transform their structures, and approaches designed to manipulate structure and functions of proteins and nucleic acids.
DNA microarray, or DNA chip technology is a new and powerful means of discovering, characterizing and analyzing genes and their expression patterns. This manual assembles and synthesizes the expertise of over 30 innovators in this emerging field to provide authoritative, detailed instruction on the design, construction, and applications of microarrays, as well as comprehensive descriptions of the software tools and strategies required for extensive image and data analysis. It complements the best selling manual "Molecular Cloning".
There are two books in this series. Volume 1 concentrates on the procedures for DNA and RNA manipulation, purification, electrophoresis and the construction and cloning of recombinant molecules. It also includes a survey of cloning vectors of Escherichia Coli. As with the first edition, the objective is to combine solid practical information with sufficient background material to meet the requirements of any enduser
In Volume 2 of Essential Molecular Biology, procedures for preparing gene libraries and identifying genes are described, together with methods for studying the structure of a cloned gene and the way it is expressed in the cell.
Protein expression is an increasingly important tool for research on gene function. What is needed is not just a lab manual providing established methods as well as the latest state-of-the-art protocols, but also clear advice on what expression system to choose when. This book covers expression across a broad range of systems, including the following. *Baculovirus expression vectors, *CHO cells, * E. coli, *HEK293-EBNA1 cells, * Lactococcus lactis, * S. cerevisiae, *transfected insect cells, * Pichia pastoris, *mammalian cells using BacMam viruses, *lentiviral vectors, *wheat germ cell-free system. The book takes the reader through how to make an informed choice of appropriate system, taking into account the protein target, the time involved, the ultimate use of the expressed protein, and the laboratory equipment required. In addition, the book describes the optimisation of expression strategies, expression engineering using ribosome display, and how to select protein variants with improved expression.
Leading scientists in gene expression methodology and bioinformatics data analysis describe readily reproducible methods for measuring RNA levels in cells and tissues. The techniques presented include new methods for applying the Affymetrix GeneChip, SAR-SAGE, StaRT-PCR, SSH, the Invader Assay, and ADGEM. The authors also provide critical bioinformatics insight and resources for data analysis and management. By distilling the basic underlying principles of many methods to a few straightforward concepts, investigators can easily choose the method most appropriate to their application
This sixth edition of James D. Watson′s classic textbook Molecular Biology of the Gene has been thoroughly revised and updated. Accessible to anyone interested in molecular biology and genetics, the book provides a historical basis for the field, concise descriptions of fundamental chemical concepts, a comprehensive survey of genome maintenance and expression, and a discussion of standard techniques and model organisms commonly used in molecular biology studies. It includes all new chapters on the regulatory RNAs and genomics and systems biology. The book has an accompanying Web site (www.aw-bc.com/watson/), which contains interactive tutorials, animations, and critical-thinking exercises designed to help students explore and visualize complex concepts.
This edition update and expands Bruce White′s best-selling "PCR 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 PCR and GC-rich template amplification. Powerful applications of PCR in library construction and sublibrary generation and screening are presented.
Using Whole Genome Amplification (WGA) methods, it is possible to create microgram quantities of DNA starting with as little as one nanogram of genomic DNA and in some cases even a single eukaryotic or bacterial cell. The implementation of such WGA methods provides an ample supply of DNA for large-scale genetic studies. This title provides a comprehensive overview of the field and will be welcomed by all researchers looking to take advantage of the latest developments.
This volume, part of the Advances in Molecular Biology series, presents detailed protocols and trouble-shooting advice on the yeast two-hybrid system, one of the most powerful and versatile methods for characterizing a protein′s functions. Topics include: characterizing hormone/receptor complexes, identifying peptide ligands, how to identify mutations that disrupt an interaction, how to construct an activation domain hybrid library, and how to dissect the cell cycle and other complex genetic networks.