PCR Cloning Protocols

PCR Cloning Protocols

Annotation 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: Springer Science & Business Media

ISBN: 9781592591770

Category: Science

Page: 439

View: 747

Annotation 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.
Categories: Science

PCR Cloning Protocols

PCR Cloning Protocols

This book provides extensive protocols on the use of PCR for molecular cloning, including the cloning of known sequences, cloning of unknown sequences and cloning of gene family members.

Author: Bruce Alan White

Publisher: Springer Science & Business Media

ISBN: 0896033430

Category: Science

Page: 490

View: 140

This book provides extensive protocols on the use of PCR for molecular cloning, including the cloning of known sequences, cloning of unknown sequences and cloning of gene family members.
Categories: Science

PCR Cloning Protocols

PCR Cloning Protocols

The techniques bring to both new and established researchers the power to apply PCR-based methodology to the cloning and modification of DNA, either through innovative protocols or by fostering individual creativity to modify and customize ...

Author: Bruce A. White

Publisher:

ISBN: OCLC:748540457

Category:

Page:

View: 738

Distinguished scientists and researchers present a comprehensive collection of current preparative PCR techniques that can be used in cloning and modifying DNA and cDNA. Topics include performing and optimizing PCR (including long PCR), cloning PCR products, cloning unknown neighboring DNA, and library construction and screening. Also covered are mutagenesis, recombination, and in vitro selection, differential and subtractive approaches to cDNA analysis and screening, and cloning members of gene families. The techniques bring to both new and established researchers the power to apply PCR-based methodology to the cloning and modification of DNA, either through innovative protocols or by fostering individual creativity to modify and customize the protocols to best fit their own needs.
Categories:

PCR Cloning Protocols

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: 0896039730

Category: Science

Page: 439

View: 935

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.
Categories: Science

PCR Protocols

PCR Protocols

Along with the cutting-edge methodologies, this volume describes many core applications, such as PCR cloning and sequencing, expression, copy number or methylation profile analysis, ‘DNA fingerprinting’, diagnostics, protein engineering ...

Author: Daniel J. Park

Publisher: Humana Press

ISBN: 1627038191

Category: Medical

Page: 348

View: 599

Known for flexibility and robustness, PCR techniques continue to improve through numerous developments, including the identification of thermostable DNA polymerases which exhibit a range of properties to suit given applications. PCR Protocols, Third Edition selects recently developed tools and tricks, contributed by field-leading authors, for the significant value that they add to more generally established methods. Along with the cutting-edge methodologies, this volume describes many core applications, such as PCR cloning and sequencing, expression, copy number or methylation profile analysis, ‘DNA fingerprinting’, diagnostics, protein engineering, interaction screening as well as a chapter highlighting workflow considerations and contamination control, crucial for all PCR methods. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary reagents and materials, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, PCR Protocols, Third Edition seeks to further elucidate this essential technique while also providing core principles with broad applications for scientists of all backgrounds.
Categories: Medical

PCR Protocols

PCR Protocols

The correct procedures you need for frustration-free PCR methods and applications are contained in this complete, step-by-step, clearly written, inexpensive manual.

Author: Michael A. Innis

Publisher: Academic Press

ISBN: 9780080886718

Category: Science

Page: 482

View: 313

The correct procedures you need for frustration-free PCR methods and applications are contained in this complete, step-by-step, clearly written, inexpensive manual. Avoid contamination--with specific instructions on setting up your lab Avoid cumbersome molecular biological techniques Discover new applications
Categories: Science

PCR Protocols

PCR Protocols

This new volume on PCR Protocols does not attempt the impossible task of representing all PCR-based protocols.

Author: Bruce Alan White

Publisher: Springer Science & Business Media

ISBN: 9781592595020

Category: Science

Page: 392

View: 315

PCR has been successfully utilized in every facet of basic, cli- cal, and applied studies of the life sciences, and the impact that PCR has had on life science research is already staggering. C- comitant with the essentially universal use of PCR has been the creative and explosive development of a wide range of PCR-based techniques and applications. These increasingly numerous pro- cols have each had the general effect of facilitating and acceler- ing research. Because PCR technology is relatively easy and inexpensive, PCR applications are well within the reach of every research lab. In this sense, PCR has become the "equalizer" between "small" and "big" labs, since its use makes certain projects, especially those related to molecular cloning, now far more feasible for the small lab with a modest budget. This new volume on PCR Protocols does not attempt the impossible task of representing all PCR-based protocols. Rather, it presents a range of protocols, both analytical and preparative, that provide a solid base of knowledge on the use of PCR in many c- mon research problems. The first six chapters provide some basic information on how to get started. Chapters 7-19 represent primarily analytical uses of PCR, both for simple DNA and RNA detection, as well as for more complex analyses of nucleic acid (e. g. , DNA footprin ting, RNA splice site localization). The remaining chapters represent "synthetic," or preparative, uses of PCR.
Categories: Science

Neuropeptide Y Protocols

Neuropeptide Y Protocols

In Neuropeptide Y Protocols, a strong cohort of established investigators from a variety of disciplines detail their best techniques for advancing NPY research.

Author: Ambikaipakan Balasubramaniam

Publisher: Springer Science & Business Media

ISBN: 9781592590421

Category: Medical

Page: 256

View: 884

It is now clear that neuropeptide Y (NPY), the most abundant peptide present in the mammalian nervous system, is implicated in the pathophysiology of many diseases, including eating disorders, seizures, memory loss, anxiety, depression, and cardiovascular diseases, and offers major therapeutic opportunities for their treatment. In Neuropeptide Y Protocols, a strong cohort of established investigators from a variety of disciplines detail their best techniques for advancing NPY research. Among the many readily reproducible methods described are those for cloning, for the production of transgenic and knockout models, and for the use of antisense technology to define the receptors and the signal transduction pathways mediating NPY actions. A range of the major qualitative and quantitative techniques used to study tissue mRNA distribution are also included, as well as methods for radioligand binding. The cloning techniques utilize genomic DNA isolation, expression cloning, PCR cloning, the construction of hybrid receptors, and homology-based cloning. Highly practical and state-of-the-art, Neuropeptide Y Protocols constitutes a gold-standard collection of proven techniques for advancing today's understanding of this significant neuropeptide, as well as for developing the promising NPY-based therapeutics now emerging for many important disorders.
Categories: Medical

The Condensed Protocols from Molecular Cloning a Laboratory Manual

The Condensed Protocols from Molecular Cloning   a Laboratory Manual

The Condensed Protocols From Molecular Cloning: A Laboratory Manualis a single–volume adaptation of the three–volume third edition of Molecular Cloning: A Laboratory Manual.This condensed book contains only the ...

Author: Joseph Sambrook

Publisher: CSHL Press

ISBN: 9780879697716

Category: Science

Page: 800

View: 286

The Condensed Protocols From Molecular Cloning: A Laboratory Manualis a single–volume adaptation of the three–volume third edition of Molecular Cloning: A Laboratory Manual.This condensed book contains only the step–by–step portions of the protocols, accompanied by selected appendices from the world's best–selling manual of molecular biology techniques. Each protocol is cross–referenced to the appropriate pages in the original manual. This affordable companion volume, designed for bench use, offers individual investigators the opportunity to have their own personal collection of short protocols from the essential Molecular Cloning.
Categories: Science

Positional Cloning by Exon Trapping and cDNA Selection

Positional Cloning by Exon Trapping and cDNA Selection

This comprehensive guide details the exon-trapping and cDNA selection processes step by step-from isolating genomic templates and nuclear splicing, to verifying generated clones and sequenced data, to analyzing exon libraries and cDNA ...

Author: Bernhard Korn

Publisher: John Wiley & Sons

ISBN: 0471297976

Category: Science

Page: 77

View: 617

Tremendous strides in decoding the human genome have been made over the last ten years. Large, chromosome-scale, genome-scale EST-sequencing and mapping techniques have dramatically expanded the base of identified genes and sequenced DNA. Gene isolation and cloning techniques that emerged earlier on, however, still remain effective mainstays for constructing dense transcript maps to isolate coding sequences and disease-associated genes contained within a specific chromosomal region. Positional Cloning by Exon Trapping and cDNA Selection examines two powerful methods for locating the coding region of a given gene by isolating gene fragments from individual clones or pools of genomic clones. This comprehensive guide details the exon-trapping and cDNA selection processes step by step-from isolating genomic templates and nuclear splicing, to verifying generated clones and sequenced data, to analyzing exon libraries and cDNA sublibraries. Procedures covered include: * Exon-trapping systems and descriptions of pSPL1 and pSPL3 vectors. * Exon amplification protocols for preparing vectors for cloning; subcloning genomic DNA into vectors; transformation, analysis, and transfection of sublibraries; RNA transcription; and PCR amplification and PCR product cloning. * Evaluation of exon libraries by PCR colony testing, identifying artifactual clones using Southern blot, and sequencing and mapping back candidate exons. * Isolation of genomic templates, such as COSMID-, P1-, PAC, and YAC DNA. * cDNA selection experiment protocols including cDNA screening, hybridization, and biotinylation; preparation of genomic and cDNA sources; and PCR cloning. * Clone analysis and analysis by hybridization. Positional Cloning by Exon Trapping and cDNA Selection offers researchers, scientists, and graduate students an invaluable tool for probing gene distribution and molecular organization. Most importantly, it provides a critical approach to isolating specific disease genes within a targeted genomic area.
Categories: Science