Recombinant DNA methods are powerful, revolutionary techniques that allow the isolation of single genes in large amounts from a pool of thousands or millions of genes and the modification of these isolated genes or their regulatory regions for reintroduction into cells for expression at the RNA or protein levels. These attributes lead to the solution of complex biological problems and the production of new and better products in the areas of medicine, agriculture, and industry. Recombinant DNA Methodology, a volume in the Selected Methods in Enzymology series produced in benchtop format, contains a selection of key articles from Volumes 68, 100, 101, 153, 154, and 155 of Methods in Enzymology. The essential and widely used procedures provided at an affordable price will be an invaluable aid to the graduate student and the researcher. Enzymes in DNA research DNA isolation, hybridization, and cloning DNA sequence analysis cDNA cloning Gene products Identification of cloned genes and mapping of genes Monitoring cloned gene expression Cloning and transferring of genes into yeast cells Cloning and transferring of genes into plant cells Cloning and transferring of genes into animal cells Site-directed mutagenesis Protein engineering Expression vectors

Enzymes in recombinant DNA research. DNA-joining enzymes: a review. Guide to the use of type II restriction endonucleases. Site-specific cleavage of DNA at 8-,9-, and 10-bp sequences. Exonuclease III: use for DNA sequence analysis and in specific deletions of nucleotides. Methods for isolation, purification, or amplification of DNA. Gel electrophoresis of restriction fragments. Purification, specific fragmentation, and separation of large DNA molecules. Orthogonal-field-alternation gel electrophoresis. A rapid alkaline extraction method for the isolation of plasmid DNA. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Vectors or methods for gene cloning. Escherichia coli plasmids packageable in vitro in a bacteriophage particles. Production of single-stranded plasmid DNA. High-efficiency cloning of full-length cDNA: construction and screening of cDNA expression libraries for mannalian cells. Transformation and preservation of competent bacterial cells by freezing. Plasmid screening high colony density. New bacteriophage lambda vectors with positive selection for cloned inserts. A phage vectors-EMBL series. Agt 11: gene isolation with antibody probes and other applications. One-step gene disruption in yeast. Cloning regulated yeast genes from a pool of lacZ fusions. Selection procedure for isolation fo centromere DNAs from rom saccharomyces cerevisiae. Construction of high copy yeast vectors using 2-um circle sequences. Improved vectors for plant transformation: expression cassette ...

Enzymes are indispensable tools in recombinant DNA technology and genetic engineering. This book not only provides information for enzymologists, but does so in a manner that will also aid nonenymologists in making proper use of these biocatalysts in their research. The Enzymology Primer for Recombinant DNA Technology includes information not usually found in the brief descriptions given in most books on recombinant DNA methodology and gene cloning. Provides essential basics as well as up-to-date information on enzymes most commonly used in recombinant DNA technology Presents information in an easily accessible format to serve as a quick reference source Leads to a better understanding of the role of biocatalysts in recombinant DNA techniques

The critically acclaimed laboratory standard for forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerlyawaited, frequently consulted, and praised by researchers and reviewers alike. More than 250 volumes have been published (all of them still in print) and much of the material is relevant even today--truly an essential publication for researchers in all fields of life sciences.

The critically acclaimed laboratory standard for forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerlyawaited, frequently consulted, and praised by researchers and reviewers alike. More than 250 volumes have been published (all of them still in print) and much of the material is relevant even today--truly an essential publication for researchers in all fields of life sciences. * Methods for: * DNA isolation and cloning* Synthesizing complementary DNA (cDNA)* Cleaving and manipulating DNA * Selecting useful reporter genes* Constructing vectors for cloning genes* Constructing expression vectors* Site-directed mutagenesis and gene disruption* Identifying and mapping genes* Transforming animal and plant cells* Sequencing DNA* Amplifying and manipulating DNA and PCR* Detecting DNA - protein interaction

This comprehensive yet balanced work emphasizes the principles and rationale underlying recombinant DNA methodology while furnishing a general understanding of the experimental protocols-suggesting flexible approaches to resolving particular molecular necessities that are easily adaptable to readers' specific applications. Features summary tables presenting at-a-glance information on practices of recombinant DNA methodologies! Recombinant DNA Principles and Methodologies discusses basic and advanced topics requisite to the employment of recombinant DNA technology, such as plasmid biology nucleic acid biochemistry restriction enzymes cloning strategies gel electrophoresis southern and northern blotting preparation of probes phage lambda biology cosmids and genome analysis cloned gene expression polymerase chain reaction conventional and automated DNA sequencing site-directed mutagenesis and more! Elucidating the material with over 2250 edifying references, equations, drawings, and photographs, this state-of-the-art resource is a valuable hands-on guide for molecular and cell biologists, biochemists, bioprocess technologists, applied and industrial microbiologists, virologists, geneticists, chemical engineers, and upper-level undergraduate and graduate students in these disciplines.

Calculations for Molecular Biology and Biotechnology: A Guide to Mathematics in the Laboratory, Second Edition, provides an introduction to the myriad of laboratory calculations used in molecular biology and biotechnology. The book begins by discussing the use of scientific notation and metric prefixes, which require the use of exponents and an understanding of significant digits. It explains the mathematics involved in making solutions; the characteristics of cell growth; the multiplicity of infection; and the quantification of nucleic acids. It includes chapters that deal with the mathematics involved in the use of radioisotopes in nucleic acid research; the synthesis of oligonucleotides; the polymerase chain reaction (PCR) method; and the development of recombinant DNA technology. Protein quantification and the assessment of protein activity are also discussed, along with the centrifugation method and applications of PCR in forensics and paternity testing. Topics range from basic scientific notations to complex subjects like nucleic acid chemistry and recombinant DNA technology Each chapter includes a brief explanation of the concept and covers necessary definitions, theory and rationale for each type of calculation Recent applications of the procedures and computations in clinical, academic, industrial and basic research laboratories are cited throughout the text New to this Edition: Updated and increased coverage of real time PCR and the mathematics used to measure gene expression More sample problems in every chapter for readers to practice concepts