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Molecular Analysis And Genome Discovery Ralph Rapley



Molecular Analysis and Genome Discovery, Second Edition is a completely revised and updated new edition of this successful book. The text provides a comprehensive overview of recent developments in the fast moving field of molecular based diagnostics of disease markers. Key concepts and applications are provided alongside practical information on current techniques currently being researched and developed. Each chapter offers an up-to-date analysis of the subject encompassing the very latest technology platforms and is an essential reference for researchers in the field looking for an up-to-date overview of the subject. The book will also be an indispensable resource for those working in the biotechnology and pharmaceutical industries. New for this edition: chapters on Genotyping through Mutation Detection; Differential Gene Expression; Haplotyping and Molecular Profiling.




Molecular Analysis and Genome Discovery Ralph Rapley



SNPs, dichotomous (biallelic) markers, have been developed in many species [e.g. [20, 21]] including grape where they were derived from BAC and EST libraries and used successfully to build genetic maps [22, 23] and to anchor them to a physical map [24]. In addition, SNPs identified in grape gene sequences have been employed in genetic diversity studies [25, 26] and linkage disequilibrium analyses [27, 28]. Moreover, the recent decoding of the grape genome sequence in the heterozygous Pinot Noir cultivar provided the grape research community with 1,700,000 SNPs from coding and non-coding regions [6]. Different strategies have been applied in grape for SNP detection and genotyping, including low to mid [29] and high throughput [30] methods. However, most of the SNP discovery and application has been limited to V. vinifera. The use and transferability of SNPs across V. vinifera has been restricted to a few cultivars [23] and to a few wild forms [27]. In addition to the development of markers for MAS, knowledge on transferability of SNPs is required to allow the identification of useful alleles for diversity and association studies. To date, a comprehensive study of SNP use and transferability across species within Vitis has never been attempted.


This study was intended to validate the use of 137 SNP markers, developed from the heterozygous genome of Pinot Noir clone ENTAV 115. The transferability of these markers was assessed across V. vinifera cultivars, wild forms of V. vinifera, and non-vinifera Vitis species to validate their utility as informative tools for marker-assisted selection in grape improvement programs, diversity studies, association analysis and mapping purposes.


Advances in molecular biology and biotechnology are increasing at a rapid pace, both in the development of new methodologies and in their practical applications. This popular textbook has been revised and updated to provide an overview of this exciting area of bioscience and to reflect a number of the key developments driving this expansion. Chapters on the basic methods of key technologies such as nucleic acid analysis and bioinformatics are presented, in addition to genomics and proteomics, which highlight the impact of molecular biology and biotechnology. New chapters on important and emerging methods have been introduced such as gene editing, next generation sequencing, nanobiotechnology and molecular modelling.


The first six chapters deal with the core technology used in current molecular biology and biotechnology. These primarily deal with basic molecular biology methods such as PCR, cloning genes and genomes, protein analysis techniques and recombinant protein production. Later chapters address major advances in the applications of specialist areas of molecular biotechnology. Experienced lecturers and researchers have written each chapter and the information is presented in an easily assimilated form. This book makes an ideal text for undergraduates studying these areas and will be of particular interest to students in many areas of biosciences, biology and chemistry. In addition, it will appeal to postgraduates and other scientific workers who need a sound introduction to this ever rapidly advancing and expanding area.


The field of molecular biology and biotechnology continues to be of major importance and underpins nearly all areas of the biosciences; indeed, the advances in many fields have continued at a spectacular pace with an ever-accelerating rate of progress. Since the 6th edition of this book was published, significant developments in many areas of bioscience have taken place. Fields such as gene editing and genome analysis, vaccine development, nanobiotechnology and antibody engineering have all seen major refinements, giving rise to the development of new methodologies and a focus on new applications. This has provided further understanding and characterisation of biochemical pathways and cellular interactions and has accelerated the molecular understanding of plant, virus and human processes in health and disease.


The 7th edition of this book has been updated and augmented with new chapters to provide an overview of these important areas. The early chapters deal with core technologies in the biosciences, including basic molecular biology, genes and genomes, expression of recombinant proteins, proteomics and transgenesis. A series of chapters then deal with more specialised topics, reflecting current challenges in biotechnology, including molecular analysis of yeast, antibody engineering and immunotherapeutics and human and animal cell culture.


Genome editing and genome sequencing are new chapters that address the unparalleled advances seen in these areas in recent years. In addition, further new chapters on bioinformatics, nanobiotechnology, molecular modelling and intellectual property have been introduced and chapters on agricultural biotechnology, biosensors and vaccines have been updated for this new edition. It is the intention that the coverage of many of the key areas in molecular biotechnology will serve as a solid foundation for those embarking on, or engaged in, studies of these exciting fields. As such, this 7th edition of Molecular Biology and Biotechnology will be of particular interest to undergraduate students in the biosciences, biotechnology and chemistry, and also to postgraduates and other scientists who require a sound introduction to this rapidly advancing and expanding area.


When little DNA information is available to prepare a gene probe, it is possible in some cases to use the knowledge gained from analysis of the corresponding protein. Thus it is possible to isolate and purify proteins and sequence part of the N-terminal end of the protein. From our knowledge of the genetic code, it is possible to predict the various DNA sequences that could code for the protein and then synthesise appropriate oligonucleotide sequences chemically. Owing to the degeneracy of the genetic code, most amino acids are coded for by more than one codon, hence there will be more than one possible nucleotide sequence that could code for a given polypeptide. The longer the polypeptide, the greater is the number of possible oligonucleotides that must be synthesised. Fortunately, there is no need to synthesise a sequence longer than about 20 bases, since this should hybridise efficiently with any complementary sequences and should be specific for one gene. Ideally, a section of the protein should be chosen that contains as many tryptophan and methionine residues as possible, since these have unique codons and there will therefore be fewer possible base sequences that could code for that part of the protein. The synthetic oligonucleotides can then be used as probes in a number of molecular biology methods. Indeed, the chemical solid-phase synthesis of DNA for probe or gene synthesis has improved over the years since it was first introduced and limits on length, construction and error correction or fidelity have also been developed. Recent advances have investigated enzymatic synthesis using terminal transferase (TdT) and new formats termed DNA printing. Refinement of these exciting methods may eventually bring DNA synthesis to benchtop computer-controlled machines. 2ff7e9595c


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