PRINS and In Situ PCR Protocols

PRINS and In Situ PCR Protocols
Author: Franck Pellestor
Publisher: Springer Science & Business Media
Total Pages: 257
Release: 2008-02-03
Genre: Science
ISBN: 1597450685

The in situ hybridization and PCR technologies are now well-established molecular techniques for studying chromosomal aneuploidy and rearran- ments, gene localization and expression, and genomic organization. Over the last decade, we have seen increasing applications in these fields. By combining the high sensitivity of the PCR reaction and the cytological localization of target sequences, both PRINS and in situ PCR techniques have provided highly powerful complements to FISH for in situ cellular and molecular investigations. Both these approaches have several advantages in terms of sensitivity and specificity, owing to the use of primers and to the fast kinetics of annealing and elongation reactions in situ. In the first edition of PRINS and In Situ PCR Protocols edited by John R. Gosden, experts in the field presented in detail a variety of applications of PRINS and in situ PCR techniques, in a wide range of clinical conditions. Since the publication of this successful reference book, there have been s- nificant improvements in in situ detection techniques. This completely revised and updated second edition presents a compreh- sive selection of new procedures developed in the field of PRINS and in situ PCR technologies. The book has two sections. Part I, Basic Methodology, contains chapters that provide useful protocols for many variations of PRINS and in situ PCR, including a new fast multicolor PRINS method, and protocols for PRINS detection of unique sequences in situ.

PCR Protocols

PCR Protocols
Author: John M. S. Bartlett
Publisher: Springer Science & Business Media
Total Pages: 1083
Release: 2008-02-03
Genre: Science
ISBN: 1592593844

In this new edition, the editors have thoroughly updated and dramatically expanded the number of protocols to take advantage of the newest technologies used in all branches of research and clinical medicine today. These proven methods include real time PCR, SNP analysis, nested PCR, direct PCR, and long range PCR. Among the highlights are chapters on genome profiling by SAGE, differential display and chip technologies, the amplification of whole genome DNA by random degenerate oligonucleotide PCR, and the refinement of PCR methods for the analysis of fragmented DNA from fixed tissues. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes a background introduction outlining the principle behind the technique, equipment and reagent lists, tips on trouble shooting and avoiding known pitfalls, and, where needed, a discussion of the interpretation and use of results.

In Situ Hybridization Protocols

In Situ Hybridization Protocols
Author: Ian A. Darby
Publisher: Taylor & Francis US
Total Pages: 368
Release: 2000
Genre: Medical
ISBN: 9780896036864

Annotation Darby (human biology, RMIT U., Victoria, Australia) is joined by geneticists, molecular biologists, and pathologists from around the world to describe basic and advanced techniques for hybridization, for whole-mount embryo specimens and at the electron microscope level. Coverage includes protocols for detection of DNA fragmentation in apoptosis, localization of genes to particular chromosomes, and the use of DNA and RNA probes to detect expression in cells or tissue sections. For novice and experienced investigators who need proven and readily reproducible methods. Annotation c. Book News, Inc., Portland, OR (booknews.com)

In-Situ PCR Techniques

In-Situ PCR Techniques
Author: Omar Bagasra
Publisher: Wiley-Liss
Total Pages: 0
Release: 1997-07-04
Genre: Science
ISBN: 9780471159469

This book describes comprehensive step-by-step protocols for the delineation of genetic amplification and histological detection techniques. Each procedure has been tested and validated for its sensitivity, precision, and reproducibility, and the authors give advice on the design of primers for PCR applications and on optimizing these protocols for use with plant, insect, and prokaryotic cells.

PRINS and PNA Technologies in Chromosomal Investigations

PRINS and PNA Technologies in Chromosomal Investigations
Author: Franck Pellestor
Publisher: Nova Publishers
Total Pages: 208
Release: 2007
Genre: Science
ISBN: 9781600214134

Book & CD. Advances in molecular biotechnology have greatly improved the sensitivity and the efficiency of methods utilised for genetic investigations and diagnosis. In the domain of chromosome analysis, the introduction of molecular techniques has led to the development of a new approach, called Molecular Cytogenetics, which has surpassed previously available techniques to become a foremost biological method. The fluorescence in situ hybridisation (FISH) is quickly became the standard technique for in situ chromosomal investigations, as illustrated by its large variety of applications in research and diagnosis. However, during the last decade, alternative methods to FISH have been introduced and have shown to be valuable in detecting chromosomes and quantifying chromosomal abnormalities. These alternative procedures are the Primed IN Situ (PRINS) labelling and the Peptide Nucleic Acid (PNA) probes. The two procedures present several advantages for the in situ detection of nucleic acid sequences, such as the small size of PNA probes and PRINS primers, or the fast kinetics of PRINS and PNA labelling reactions, that make them very attractive for a number of cytogenetic purposes. This book provides a valuable introduction and overview of the principles and the applications of alternative approaches in the field of molecular cytogenetics.

Immunochemical Protocols

Immunochemical Protocols
Author: John Pound
Publisher: Springer Science & Business Media
Total Pages: 501
Release: 2008-02-03
Genre: Science
ISBN: 1592592570

This much anticipated second edition provides a user-friendly, up-to-date handbook of reliable immunochemical techniques optimized for molecular biologists. It covers the breadth of relevant established methods from protein blotting and immunoassays through to visualization of cellular antigens and in situ hybridization, each with their latest refinements. Protocols for the production and purification of important classes of immunochemical reagents are also provided, including "conventional" and recombinant antibodies, fusion proteins and their various conjugates. This book will open the door to a new generation of immunochemical reagents with exciting possibilities.

Transmembrane Signaling Protocols

Transmembrane Signaling Protocols
Author: Dafna Bar-Sagi
Publisher: Springer Science & Business Media
Total Pages: 631
Release: 1998
Genre: Cell membranes
ISBN: 0896034887

This collection of practical, cutting-edge techniques for the study of cell signaling provides detailed, step-by-step instructions, helpful notes, and troubleshooting tips that make even the most powerful of the newest techniques readily reproducible. The protocols presented include the use of peptide libraries to study transmembrane signaling; the use of single-cell assays to analyze signal transduction pathways; the reconstitution of signaling complexes; methods for analyzing protein-protein interactions, and more. Introductory reviews explain the basic theory and enable researchers new to the area to rapidly gain understanding, as well as command of the practical knowledge and expertise afforded by the protocols. Transmembrane Signaling Protocols makes available to all researchers the many state-of-the-art techniques that have recently led to landmark discoveries in transmembrane signaling.

Molecular Biomethods Handbook

Molecular Biomethods Handbook
Author: Ralph Rapley
Publisher: Springer
Total Pages: 708
Release: 2007-10-09
Genre: Science
ISBN: 1592596428

An authoritative team of investigators illuminate the core bioanalytical techniques used every day in their own laboratories, and laboratories throughout the world. These highly experienced scientists fully explain both the theory behind, and the application of, these key techniques, and include extensive references for those seeking detailed laboratory protocols. The techniques covered range from the extraction, separation, detection, and characterization of nucleic acids to gene cloning and library production, mapping, expression, transgenesis, differential display, and DNA profiling, to name a few. Numerous key protein methods, as well as support and related techniques, are also included. The goal is to provide established scientists and novices who are new to these techniques with a deeper understanding of the widest variety of biotechniques and their applications.

Hormone Assays in Biological Fluids

Hormone Assays in Biological Fluids
Author: Michael J. Wheeler
Publisher: Springer Science & Business Media
Total Pages: 259
Release: 2008-02-04
Genre: Science
ISBN: 1592599869

Expert researchers who have developed and applied significant new assays describe in step-by-step detail a variety of methods for measuring a broad variety of hormones, related peptides, and synthetic steroids in various biological fluids. The hormones measured range from glucocorticoids in biological fluids, urinary steroids, aldosterone in blood, and plasma renin activity, to gut hormones in plasma, melatonin, prolactin, 6-sulfatoxymelatonin, and androgens in blood, saliva, and hair. The emphasis is on noncommercial assays so that investigators can set up novel methods suited to their special needs. Commercial assays are also described for comparative purposes. Tutorials on radioimmunoassay, gas chromatography-mass spectrometry, high-performance liquid chromatography, and PCR techniques help the reader to choose the best method for his or her purpose.

Differential Display Methods and Protocols

Differential Display Methods and Protocols
Author: Peng Liang
Publisher: Springer Science & Business Media
Total Pages: 321
Release: 2008-02-04
Genre: Science
ISBN: 1592599680

Since the first edition of this book dedicated to differential display (DD) technology was published in 1997, we have witnessed an explosive interest in studying differential gene expression. The gene-hunting euphoria was initially powered by the invention of DD, which was gradually overtaken by DNA microarray technology in recent years. Then why is there still the need for second edition of this DD book? First of all, DD still enjoys a substantial lead over DNA microarrays in the ISI citation data (see Table 1), despite the h- dreds of millions of dollars spent each year on arrays. This may come as a surprise to many, but to us it implies that many of the DNA microarray studies went unpublished owing to their unfulfilled promises (1). Second, unlike DNA microarrays, DD is an “open”-ended gene discovery method that does not depend on prior genome sequence information of the organism being studied. As such, DD is applicable to the study of all living organisms—from bacteria, fungi, insects, fish, plants, to mammals—even when their genomes are not sequenced. Second, DD is more accessible technically and financially to most cost-conscious “cottage-industry” academic laboratories. So clearly DD still has its unique place in the modern molecular biological toolbox for gene expression analysis.