In Vitro Transcription And Translation Protocols

Author : Martin J. Tymms
Publisher : Springer Science & Business Media
Page : 426 pages
File Size : 53,7 Mb
Release : 2008-02-02
Category : Science
ISBN : 9781592595242

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In Vitro Transcription and Translation Protocols by Martin J. Tymms Pdf

Most laboratories conducting studies that use molecular biology techniques employ in vitro transcription and translation systems as a routine part of their day-to-day research. The commercial availability of purified bacterial RNA polymerase and the availability of robust tra- lation systems has made in vitro systems attractive not only as an alt- native to the in vivo expression of genes, but also as good model systems for studying specific aspects of transcription and translation. Although fairly efficient eukaryotic translation systems have been established for a number of years, reconstitution of transcription in vitro has proved to be more difficult. Recent improvements in fractionation techniques and the cloning of proteins involved in transcription have made this a fast moving area of research. Considerable progress has also been made in recent years in developing in vitro systems to study transcription and translation in chloroplasts and mitochondria, together with systems for the study of protein import. In Vitro Transcription and Translation Protocols provides many detailed experimental procedures for prokaryotic transcription and translation systems, together with protocols for many key techniques used in the analysis of eukaryotic transcription. In keeping with the successful format of preceding volumes of the Methods in Molecular Biology series, step-by-step instructions are provided, together with extensive notes that cover troubleshooting and special tips considered important.

Author : Guido Grandi
Publisher : Springer Science & Business Media
Page : 568 pages
File Size : 46,6 Mb
Release : 2007-05-03
Category : Medical
ISBN : 9781588295583

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In Vitro Transcription and Translation Protocols by Guido Grandi Pdf

This book is a highly anticipated update of the previous edition. It provides molecular biology laboratories with the most powerful techniques for exploiting in vitro transcription and translation systems. It has been completely updated with new chapters and topics.

Author : Guido Grandi
Publisher : Humana Press
Page : 276 pages
File Size : 55,5 Mb
Release : 2007-05-03
Category : Science
ISBN : 1588295583

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In Vitro Transcription and Translation Protocols by Guido Grandi Pdf

A highly anticipated update of the previous edition, In Vitro Transcription and Translation Protocols, Second Edition, provides molecular biology laboratories with the most powerful techniques for exploiting in vitro transcription and translation systems. Completely updated with new chapters and topics, there is in-depth analysis of current technologies and applications of in-vitro transcriptions and translations systems. Detailed protocols for protein production using different in vitro transcription and translation systems are included.

Author : F. Sierra
Publisher : Birkhäuser
Page : 145 pages
File Size : 55,6 Mb
Release : 2013-11-21
Category : Science
ISBN : 9783034863834

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A Laboratory Guide to In Vitro Transcription by F. Sierra Pdf

Author : Ralph Rapley,John M. Walker
Publisher : Springer
Page : 708 pages
File Size : 48,8 Mb
Release : 2007-10-09
Category : Science
ISBN : 9781592596423

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Molecular Biomethods Handbook by Ralph Rapley,John M. Walker Pdf

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.

Author : Jeff Braman
Publisher : Springer Science & Business Media
Page : 284 pages
File Size : 43,6 Mb
Release : 2008-02-05
Category : Science
ISBN : 9781592591947

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In Vitro Mutagenesis Protocols by Jeff Braman Pdf

Hands-on researchers with proven track records describe in stepwise fashion their advanced mutagenesis techniques. The contributors focus on improvements to conventional site-directed mutagenesis, including a chapter on chemical site-directed mutagenesis, PCR-based mutagenesis and the modifications that allow high throughput mutagenesis experiments, and mutagenesis based on gene disruption (both in vitro- and in situ-based). Additional methods are provided for in vitro gene evolution; for gene disruption based on recombination, transposon, and casette mutagenesis; and for facilitating the introduction of multiple mutations. Time-tested and highly practical, the protocols in In Vitro Mutagenesis Protocols, 2nd Edition offer today's molecular biologists reliable and powerful techniques with which to illuminate the proteome.

Author : I.M. Rosenberg
Publisher : Springer Science & Business Media
Page : 453 pages
File Size : 52,6 Mb
Release : 2013-03-14
Category : Mathematics
ISBN : 9781475711080

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Protein Analysis and Purification by I.M. Rosenberg Pdf

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.

Author : Annette M. Griffin,Hugh G. Griffin
Publisher : Springer Science & Business Media
Page : 434 pages
File Size : 43,9 Mb
Release : 2008-02-02
Category : Science
ISBN : 9781592595129

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Computer Analysis of Sequence Data Part II by Annette M. Griffin,Hugh G. Griffin Pdf

DNA sequencing has become increasingly efficient over the years, resulting in an enormous increase in the amount of data gener ated. In recent years, the focus of sequencing has shifted, from being the endpoint of a project, to being a starting point. This is especially true for such major initiatives as the human genome project, where vast tracts of DNA of unknown function are sequenced. This sheer volume of available data makes advanced computer methods essen tial to analysis, and a familiarity with computers and sequence analy sis software a vital requirement for the researcher involved with DNA sequencing. Even for nonsequencers, a familiarity with sequence analysis software can be important. For instance, gene sequences already present in the databases can be extremely useful in the design of cloning and genetic manipulation experiments. This two-part work on Computer Analysis of Sequence Data is designed to be a practical aid to the researcher who uses computers for the acquisition, storage, or analysis of nucleic acid (and/or pro tein) sequences. Each chapter is written such that a competent scien tist with basic computer literacy can carry out the procedure successfully at the first attempt by simply following the detailed prac tical instructions that have been described by the author. A Notes section, which is included at the end of each chapter, provides advice on overcoming the common problems and pitfalls sometimes encoun tered by users of the sequence analysis software.

Author : Philip Shepherd,Christopher J. Dean
Publisher : Oxford University Press, USA
Page : 520 pages
File Size : 48,5 Mb
Release : 2000
Category : Immunologic techniques
ISBN : 0199637229

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Monoclonal Antibodies by Philip Shepherd,Christopher J. Dean Pdf

Monoclonal Antibodies: A Practical Approach covers the preparation, testing, derivation, and applications of monoclonal antibodies. New immunological techniques incorporating tried and tested methodologies are described, making the book of interest to established and inexperienced immunologists. Both the standard somatic hybridization technique and recombinant techniques, including the use of phage libraries, for the preparation of rodent and human monoclonal antibodies are described. Protocols for both the small and large scale production are detailed, as well as purification and labelling (with both radioisotopes and non-radioisotopes) methods. The applications of monoclonal antibodies in immunoblotting, enzyme linked immunoassays, immunofluorescence, and FACS analysis are all covered in detail. Finally protocols are given for the use of monoclonal antibodies in rheumatoid arthritis, tissue typing, detecting DNA modified during chemotherapy, and in the clinical analysis of transplantation samples for malignancy. This book will therefore be an invaluable laboratory companion to anyone using monoclonal antibodies in their research.

Author : Kevin D. Altria
Publisher : Springer Science & Business Media
Page : 347 pages
File Size : 55,8 Mb
Release : 2008-02-02
Category : Science
ISBN : 9781592595396

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Capillary Electrophoresis Guidebook by Kevin D. Altria Pdf

This book is intended to be a working guide to the operation of capillary electrophoresis (CE) instrumentation. Since CE is still a rap idly maturing technique, detailed validated protocols are not widely established. Therefore, extensive experimental procedures are not pro vided for individual analyses. The intention is to provide general guide lines on the principles and practice of CE and to give an overview of the specific technologies and important application areas. Part I provides operating instructions for standard commercially available instruments. Guidelines are included for activities such as changing capillaries, method development, quantitative procedures, optimization of precision and sensitivity, and the validation of meth ods, fraction collection, and troubleshooting, as well as a quick guide to running a separation. The application range of CE is possibly the most diverse of all analytical techniques and ranges from large, complex macromolecules, such as proteins and nucleic acids, to small solutes, such as organic drugs and inorganic anions and cations.

Author : J.M. Walker,W. Gaastra
Publisher : Springer Science & Business Media
Page : 304 pages
File Size : 40,7 Mb
Release : 2012-12-06
Category : Science
ISBN : 9789401165631

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Techniques in Molecular Biology by J.M. Walker,W. Gaastra Pdf

The last few years have seen the rapid development of new methodology in the field of molecular biology. New techniques have been regularly introduced and the sensitivity of older techniques greatly improved upon. Developments in the field of genetic engineering in particular have contributed a wide range of new techniques. The purpose of this book therefore is to introduce the reader to a selection of the more advanced analytical and preparative techniques which the editors consider to be frequently used by research workers in the field of molecular biology. In choosing techniques for this book we have obviously had to be selective, and for the sake of brevity a knowledge of certain basic biochemical techniques and terminology has been assumed. However, since many areas of molecular biology are developing at a formidable rate and constantly generating new terminology, a glossary of terms has been included. The techniques chosen for this book are essentially based on those used in a series of workshops on 'techniques in molecular biology' that have been held at The Hatfield Polytechnic in recent years. In choosing these chapters we have taken into account many useful suggestions and observations made by participants at these workshops. Each chapter aims to describe both the theory and relevant practical details for a given technique, and to identify both the potential and limitations of the technique. Each chapter is written by authors who regularly use the technique in their own laboratories.

Author : Michael P. Starkey,Ramnath Elaswarapu
Publisher : Springer Science & Business Media
Page : 537 pages
File Size : 41,5 Mb
Release : 2008-02-03
Category : Medical
ISBN : 9781592592357

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Genomics Protocols by Michael P. Starkey,Ramnath Elaswarapu Pdf

We must unashamedly admit that a large part of the motivation for editing Genomics Protocols was selfish. The possibility of assembling in a single volume a unique and comprehensive collection of complete protocols, relevant to our work and the work of our colleagues, was too good an opportunity to miss. We are pleased to report, however, that the outcome is something of use not only to those who are experienced practitioners in the genomics field, but is also valuable to the larger community of researchers who have recognized the potential of genomics research and may themselves be beginning to explore the technologies involved. Some of the techniques described in Genomics Protocols are clearly not restricted to the genomics field; indeed, a prerequisite for many procedures in this discipline is that they require an extremely high throughput, beyond the scope of the average investigator. However, what we have endeavored here to achieve is both to compile a collection of procedures concerned with geno- scale investigations and to incorporate the key components of “bottom-up” and “top-down” approaches to gene finding. The technologies described extend from those traditionally recognized as coming under the genomics umbrella, touch on proteomics (the study of the expressed protein complement of the genome), through to early therapeutic approaches utilizing the potential of genome programs via gene therapy (Chapters 27–30).

Author : D. Lansing Taylor
Publisher : Springer Science & Business Media
Page : 437 pages
File Size : 42,8 Mb
Release : 2008-02-04
Category : Science
ISBN : 9781597452175

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High Content Screening by D. Lansing Taylor Pdf

There has always been some tension between proponents of hypothesis-driven and discovery-driven research in the broad field of life sciences. Academic research has been primarily focused on hypothesis-driven research. However, the success of the human genome project, a discovery-driven research approach, has opened the door to adding other types of discovery-driven research to a continuum of research approaches. In contrast, drug discovery research in the pharmaceutical industry has embraced discovery-driven research for many years. A good example has been the discovery of active compounds from large chemical libraries, through screening campaigns. The success of the human genome project has also demonstrated the need for both academic researchers and industrial researchers to now understand the functions of genes and gene products. The cell is the basic unit of life and it has been at the cellular level where function can be demonstrated most cost-effectively and rapidly. High content screening (HCS) was developed by Cellomics Inc. in the mid-1990s to address the need for a platform that could be used in the discovery-driven research and development required to understand the functions of genes and gene products at the level of the cell.

Author : Donna H. Wang
Publisher : Springer Science & Business Media
Page : 523 pages
File Size : 42,9 Mb
Release : 2008-02-01
Category : Medical
ISBN : 9781592590872

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Angiotensin Protocols by Donna H. Wang Pdf

A qualitative leap in the understanding of cardiovascular and n- ral regulation by the renin–angiotensin system, and of the role of this s- tem in tissue damage, has occurred as a result of the many recent advances in molecular genetic techniques. The cloning of the genes for the components of the renin–angiotensin system, the design of specific angiotensin receptor ligands, and the use of embryonic gene targeting te- niques for the creation of mutant strains have established that the renin– angiotensin system is important in blood pressure regulation, ion and fluid homeostasis, and tissue growth and remodeling Further investigation of the mechanisms by which this system p- ticipates in cardiovascular regulation may shed some light on the pat- genesis of several cardiovascular diseases, e. g. , hypertension, congestive heart failure, and chronic renal failure. Despite the promise of this system as a target for therapeutic interventions for these diseases, there are great challenges in the integration of the attempts to close the gap between the traditional literature of medicine and the explosion of information from the new technologies. This book’s title, Angiotensin Protocols, reflects the authors’ strong efforts to translate expert knowledge into easy-to-follow practice. The book opens with introductory chapters, and each specialty section provides detailed methods covering a wide variety of techniques, ranging from genetic manipulation of targeted genes to functional studies of the renin– angiotensin system.

Author : John R. Crowther
Publisher : Springer Science & Business Media
Page : 231 pages
File Size : 49,6 Mb
Release : 2008-02-02
Category : Medical
ISBN : 9781592595297

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ELISA by John R. Crowther Pdf

ELISA: Theory and Practice introduces to scientists at all levels of expertise the principles of the most commonly used assay technique known as the Enzyme Linked Immunosorbent Assay. The book provides readers with full descriptions of the basic systems that make ELISA one of the most powerful techniques in science today, and also examines in detail the data obtained by ELISA and their analysis and actual manipulation. ELISA: Theory and Practice is designed not only to train novices in the science of ELISA, but also to aid investigators experienced in any of the biological sciences in performing independently assays of antibodies and antigens. Mastery of the book's contents will allow readers to fully appreciate exactly how and why assays function, as well as permit the efficient development of individual assays that are both rapid and accurate.