PCR is now one of the most widely used of basic molecular biology techniques and is an indispensable research tool for the molecular biologist. The basic PCR technique provides the cornerstone for in vitro DNA amplification, upon which further features can be built to expand the range ofPCR-based applications. A Practical Approach Volume 2 is not a revised version of Practical Approach , but sets out to address some of the new and exciting applications of PCR including direct sequencing, mRNA quantitation and expression, genomic DNA mapping, and mutation analysis.
Sequence data--either lists of nucleotides or of amino acids--are now easily gathered using automated equipment; the real effort is involved in interpreting the data to produce predictions of protein structure or function. With the advent of worldwide computer networks, a plethora of software is now available for sequence analysis. This book describes the techniques for computer analysis of sequence data, with the emphasis on general issues rather than specific algorithms. Unlike many books on these topics, which focus on the "how-to" aspects of software packages, this one places more emphasis on the science behind the packages and on interpretation of the results.
This is a major update of the highly popular second edition, with changes in the content and organisation that reflect advances in the subject. New and expanded topics include cytoskeleton, molecular motors, bioimaging, biomembranes, cell signalling, protein structure, and enzyme regulation. As with the first two editions, the third edition of Instant Notes in Biochemistry provides the essential facts of biochemistry with detailed explanations and clear illustrations.
This two-volume set provides detailed practical guidance on all major aspects of RNA processing. Each procedure is clearly explained so that the reader can follow all of the key stages of a successful experimental investigation.
The book begins by describing in detail the rationale behind designing gene targeting vectors, including both basic replacement vectors for creating null mutations and more sophisticated two-step approaches for creating subtle mutations. It describes the latest techniques for culturing andmanipulating ES cells and a number of approaches for identifying rare targeted cell clones. Techniques for culturing and assaying for mouse and human hematopoietic progenitors in vitro and in vivo are then covered, followed by the design of, and use of, retrovirus vectors for introducing andexpressing genes in bone marrow cells. Two approaches for making ES cell chimeras are described. One chapter covers the more standard approach of ES cell injection into blastocysts as well as morula injection. Another chapter describes techniques for making aggregation chimeras and in particulara recent approach using tetraploid host embryos to produce completely ES cell-derived embryos and mice. The critical steps for obtaining germline chimeras are discussed. The book is concluded by a discussion of the many uses of enhancer, gene and promoter trap vectors, the advantages of each, anddescribes in detail the techniques for producing and analyzing "trap" vector insertions in ES cells. The protocols are written with sufficient detail for a novice in the area to use them as a manual to learn the various techniques required for producing mutant mice, beginning with the embryonicstem cells and ending up with chimeric mice for breeding.
In recent years, the quest for greater understanding of postimplantation mammalian development has led research workers to devise techniques which overcome the problems of inaccessibility posed by embryos in the uterus, and to adapt analytic techniques such as in situ hybridization andantibody labelling to the unique requirements of mammalian embryonic material. This book fulfills the need for a detailed practical instruction in these methods, providing an illustrated account of the morphological stages of postimplantation development. Research workers in mammalian embryologywill find this book invaluable both as a laboratory manual and reference work.
A practical response to the increase of interest in the fields of interferon and lymphokine research, this volume presents the techniques involved with these independent areas of research. Reflecting recent progress in these fields, contributors describe protocols for working with interferons including antibody preparation, immuno-assay, purification, expressing recombinant genes in heterologous systmes and quantitating mRNA with cDNA probes; the chapters on techniques for lymphokines address aspects unique to each different lymphokine. Readership: immunology, biology researchers, postgraduates.
Now in a new second edition, this popular book includes a wealth of practical information on the much-used techniques of gel electrophoresis. Authoritative and completely current, the volume provides readers with an up-to-date, "how-to" guide to this growing field.
