Basic Cloning Procedures

This manual introduces the reader to basic methods used in the isolation, cloning and analysis of genetic material.

Author: Valdis Berzins

Publisher: Springer

ISBN:

Category: Science

Page: 169

View: 508

This manual introduces the reader to basic methods used in the isolation, cloning and analysis of genetic material. The protocols include RT-PCR amplification, gene cloning, hybridization analysis and sequencing of nucleic acids, PCR-based site-specific mutagenesis, analysis of protein DNA-specific interaction, cell-free protein synthesis and product electrophoretic and immunological analysis. Each protocol includes short background information, a detailed description of the necessary materials, step-by-step procedures, a troubleshooting guide and useful practical hints.

Basic Cloning Procedures

The procedure described by Okayama and Berg (1982), as later modified by Gubler and Hoffman (1983), is currently the most widely used technique for cDNA cloning. In the first step, firststranded cDNA is synthesized using the classical ...

Author: Valdis Berzins

Publisher: Springer Science & Business Media

ISBN: 3642719651

Category: Science

Page: 169

View: 985

This manual introduces the reader to bacic methods used in the isolation, cloning and analysis of genetic material. The protocols include RT-PCR amplification, gene cloning, hybridization analysis and sequencing of nucleic acids, PCR-based site-specific mutagenesis, analysis of protein DNA-specific interaction, cell-free protein synthesis and product electrophoretic and immunological analysis. Each protocol includes short background information, a detailed description of the necessary materials, step-by-step procedures, a troubleshooting guide and useful practical hints.

Basic Methods in Molecular Biology

This book will be of interest to practitioners in the fields of biology and molecular genetics.

Author: Leonard Davis

Publisher: Elsevier

ISBN: 044460149X

Category: Science

Page: 399

View: 918

Basic Methods in Molecular Biology discusses the heart of the most recent revolution in biology—the development of the technology of genetics. The achievements in this field have simply changed what biologists do and, perhaps even more important, the way they think. Moreover, never before have scientists from such a broad range of disciplines rushed into such a small and slightly arcane field to learn and carry off a bit of the technology. This book comprises 21 chapters, opening with three introductory ones that discuss the basics of molecular biology; the tools of the molecular biologist; and general preparations, procedures, and considerations for use of the book. The following chapters then discuss cloning vectors and bacterial cells; preparation of DNA from eukaryotic cells; probing nucleic acids; plasmid DNA preparation; DNA restriction fragment preparation; purification of DNA; and preparation and analysis of RNA from eukaryotic cells. Other chapters cover preparation of DNA from bacteriophage clones; cloning DNA from the eukaryotic genome; subcloning into plasmids; M13 cloning and sequencing; further characterization of cloned DNA; transfection of mammalian cells in culture; protein methods; general methods; and specialized methods. This book will be of interest to practitioners in the fields of biology and molecular genetics.

DNA Cloning A Hands on Approach

This book offers step-by-step instruction on DNA cloning, defined as moving genes around plasmids, mutating genes, or mining new genes.

Author: Seok-Yong Choi

Publisher: Springer

ISBN: 9402416625

Category: Medical

Page: 131

View: 875

This book offers step-by-step instruction on DNA cloning, defined as moving genes around plasmids, mutating genes, or mining new genes. The aim is to provide those new to the field with reliable and up-to-date practical guidance while at the same time conveying the scope for creativity. After a brief synopsis of the history of cloning, the fundamentals and prerequisites are explained, covering, for example, software, vectors commonly used in the lab, appropriate choice of restriction endonucleases, the preparation of agarose gels, competent cells, and LB agar plates, and procedures to be followed upon receipt of new plasmids. The remainder of the book is devoted to the clear description of methods and individual steps in cloning. Guidance is provided on the cut and paste method, DNA sequencing, direct sequencing, primer design, PCR-based gene insertion and deletion, epitope tag insertion, the use of RACE technology, BAC recombineering, and much, much more. Sources of error and a variety of techniques that make life considerably easier when cloning are also examined in detail.

DNA Cloning and Assembly Methods

In DNA Cloning and Assembly Methods, expert researchers in the field detail many of the methods which are now commonly used for DNA cloning and make cloning procedures faster, more reliable and also suitable for high-throughput handling.

Author: Svein Valla

Publisher: Humana Press

ISBN: 9781627037631

Category: Medical

Page: 303

View: 747

In DNA Cloning and Assembly Methods, expert researchers in the field detail many of the methods which are now commonly used for DNA cloning and make cloning procedures faster, more reliable and also suitable for high-throughput handling. These include methods and protocols that are based on several mechanisms including type II and IIS restriction enzymes, single stranded annealing, sequence overlap, and recombination. With additional chapters on software programs that are suitable for primer design, a feature crucial for the functionality of the described methods. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, DNA Cloning and Assembly Methods seeks to provide scientist with a valuable and useful resource for wet lab researchers within life sciences.

Basic DNA and RNA Protocols

The collection includes not only many updated and improved classic techniques but also a powerful group of advanced methods that point to future progress among them nonisotopic DNA labeling silver staining and automatic sequencing.

Author: Adrian J. Harwood

Publisher: Humana PressInc

ISBN:

Category: Science

Page: 534

View: 330

An essential core collection of the latest molecular and genetic techniques for cloning subcloning sequencing PCR protein expression and much more. Each protocol represents a time-tested step-by-step recipe that creates an understanding of the procedure easily reproducible results and confidence that the procedure will work. The collection includes not only many updated and improved classic techniques but also a powerful group of advanced methods that point to future progress among them nonisotopic DNA labeling silver staining and automatic sequencing. This excellent bench companion will help those who need to learn for the first time how to conduct research on the molecular biology of nucleic acids or those who need to broaden their competence and laboratory skills. Even highly skilled researchers will find many time-saving techniques.

A Practical Guide to Molecular Cloning

This third edition of A Practical Guide to Molecular Cloning provides complete updates to all of the protocols, and incorporates much new material to cover a broader range of biomolecular techniques.

Author: Bernard Perbal

Publisher: Wiley-Liss

ISBN: 9780471235989

Category: Science

Page: 832

View: 618

This third edition of A Practical Guide to Molecular Cloning provides complete updates to all of the protocols, and incorporates much new material to cover a broader range of biomolecular techniques. It covers the key information required for students to perform cloning of any DNA fragment and eventually study its expression products. It also serves as an excellent comprehensive guide for any researcher using molecular techniques. Protocols in this established manual have been used successfully to characterize cellular localization, nature, and function of gene products in both prokaryotes and eukaryotes. This book is the most accessible laboratory manual for those who want to understand why techniques work and how best to apply them. Presents reliable, accurate, and clear stepwise protocols Includes all new coverage of genomics, proteomics (identification of targets and characterization of functional domains), PCR applications such as mutagenesis, and microarray technology and applications Discusses the biochemistry underlying commonly seen kits, in order that the reader will be able to use the kits to solve non–classical problems Includes new chapters on basic methods in microbiology, basic methods in cellular biology, methods for studying protein–protein interactions, SAGE, phage display, and methods for modifying the host genome Includes numerous tables and an appendix of companies and useful websites

PCR Cloning Protocols

PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis.

Author: Bing-Yuan Chen

Publisher: Humana Press

ISBN: 9781617372810

Category: Science

Page: 439

View: 263

PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning 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 distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination, and to clone the challenging uncharacterized DNA flanking a known DNA fragment.

Cloning

A A brief introduction to the methods for novel gene cloning , 33 A broad spectrum PCR method for the detection of polyomaviruses and avoidance of contamination by cloning vectors , 33 A century of ( epithelial ) transport physiology ...

Author: Stephen D. Fairbanks

Publisher: Nova Publishers

ISBN: 9781590338797

Category: Law

Page: 239

View: 423

The terms 'recombinant DNA technology', 'DNA cloning', 'molecular cloning' or 'gene cloning' all refer to the same process: the transfer of a DNA fragment of interest from one organism to a self-replicating genetic element such as a bacterial plasmid. The DNA of interest can then be propagated in a foreign host cell. This technology has been around since the 1970s, and it has become a common practice in molecular biology labs today. Reproductive cloning is a technology used to generate an animal that has the same nuclear DNA as another currently or previously existing animal. Dolly was created by reproductive cloning technology. In a process called 'somatic cell nuclear transfer' (SCNT), scientists transfer genetic material from the nucleus of a donor adult cell to an egg whose nucleus, and thus its genetic material, has been removed. The reconstructed egg containing the DNA from a donor cell must be treated with chemicals or electric current in order to stimulate cell division. Once the cloned embryo reaches a suitable stage, it is transferred to the uterus of a female host where it continues to develop until birth. Therapeutic cloning, also called "embryo cloning," is the production of human embryos for use in research. The goal of this process is not to create cloned human beings, but rather to harvest stem cells that can be used to study human development and to treat disease. Stem cells are important to biomedical researchers because they can be used to generate virtually any type of specialised cell in the human body. This new book presents an up-to-date Chronology of Cloning along with current and selected abstracts dealing with cloning as well as a guide to books on the topic. Access to the abstract and books sections is provided by title, subject and author indexes.

Basic Techniques in Biochemistry Microbiology and Molecular Biology

This book presents key methodologies, tools and databases for biochemistry, microbiology and molecular biology in simple and straightforward language.

Author: Aakanchha Jain

Publisher: Humana

ISBN: 9781493998630

Category: Medical

Page: 282

View: 750

This book presents key methodologies, tools and databases for biochemistry, microbiology and molecular biology in simple and straightforward language. Covering all aspects related to experimental principles and procedures, the protocols included here are brief and clearly defined, and include essential precautions to be taken while conducting experiments. The book is divided into two major sections: one on constructing, working with, and standard operating procedures for laboratory instruments; and one on practical procedures used in molecular biology, microbiology and biochemical analysis experiments, which are described in full. Each chapter describes both the basic theory and relevant practical details for a given experiment, and helps readers recognize both the experiment’s potential and limitations. Intended as an intensive introduction to the various tools used in molecular biology, the book covers all basic methods and equipment, including cloning, PCR, spectrophotometers, ELISA readers, sonicators, etc. As such, it offers a valuable asset for final year undergraduate (especially project) students, graduate research students, research scientists and technicians who wish to understand and employ new techniques in the field of biotechnology.