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El. knyga: E. coli Plasmid Vectors: Methods and Applications

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  • Formatas: PDF+DRM
  • Serija: Methods in Molecular Biology 235
  • Išleidimo metai: 03-Feb-2008
  • Leidėjas: Humana Press Inc.
  • Kalba: eng
  • ISBN-13: 9781592594092
Kitos knygos pagal šią temą:
E. coli Plasmid Vectors: Methods and ApplicationsDidesnis paveikslėlis
  • Formatas: PDF+DRM
  • Serija: Methods in Molecular Biology 235
  • Išleidimo metai: 03-Feb-2008
  • Leidėjas: Humana Press Inc.
  • Kalba: eng
  • ISBN-13: 9781592594092
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Researchers in various biological and medical sciences synthesize the current understanding and practice of using plasmids of Escherichia coli as the universal cloning host and for recombinant DNA manipulation. They discuss relevant aspects of plasmid biology; the development of plasmid vectors; how to choose the right vector and a suitable host strain; methods required to clone DNA into plasmid vectors, transform E. Coli , and analyze recombinant clones; protocols for constructing and screening libraries; and techniques for specialized cloning vehicles such as cosmids, bacterial artificial chromosomes, lambda-vectors, and phagemids. Annotation (c) Book News, Inc., Portland, OR (booknews.com)

A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression, and the use of reporter genes, are also described.

Recenzijos

...a scientific review and a methodological "cook-book" of high quality. It is truly a highly practical handbook, useful for all scientists working with gene cloning and manipulation." - Folia Microbiologica





"...a very useful addition to laboratories investigating plasmids per se, and for those employing plasmids as vectors for gene cloning." - Microbiology Today

The Function and Organization of Plasmids.- Choosing a Cloning Vector.-
Escherichia coli Host Strains.- Chemical Transformation of E. coli.-
Electroporation of E. coli.- DNA Transfer by Bacterial Conjugation.- Cosmid
Packaging and Infection of E. coli.- Isolation of Plasmids from E. coli by
Alkaline Lysis.- Isolation of Plasmids from E. coli by Boiling Lysis.-
High-Purity Plasmid Isolation Using Silica Oxide.- High-Throughput Plasmid
Extraction Using Microtiter Plates.- Isolation of Cosmid and BAC DNA from E.
coli.- Preparation of Single-Stranded DNA from Phagemid Vectors.- Using
Desktop Cloning Software to Plan, Track, and Evaluate Cloning Projects.-
Cloning in Plasmid Vectors.- Extraction of DNA from Agarose Gels.- Cloning
PCR Products with T-Vectors.- Construction of Genomic Libraries in
?-Vectors.- Rapid Screening of Recombinant Plasmids.- Restriction Analysis of
Recombinant Plasmids.- Screening Recombinant DNA Libraries.- Sequencing Using
Fluorescent-Labeled Nucleotides.- Site-Directed Mutagenesis Using the
Megaprimer Method.- Site-Directed Mutagenesis by Inverse PCR.- Creating
Nested DNA Deletions Using Exonuclease III.- Transposon and Transposome
Mutagenesis of Plasmids, Cosmids, and BACs.- In Vitro Transcription and
Translation.- Vectors for the Expression of Recombinant Proteins in E. coli.-
Expression of Recombinant Proteins From lac Promoters.- Plasmid-Based
Reporter Genes.- Plasmid-Based Reporter Genes.
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