Limit search to available items
Book Cover
Author Carson, Sue (Susan)

Title Manipulation and expression of recombinant DNA : a laboratory manual / Susan Carson, Dominique Robertson
Edition 2nd ed
Published Burlington, MA : Elsevier Academic Press, ©2006


Description 1 online resource (xvi, 151 pages) : illustrations (some color)
Contents Cover; Contents; Preface; Acknowledgments; Note to Instructors; Instrumentation; Nomenclature; Introduction; Conceptual Outline for Experiments; Experimental Procedures; Laboratory Safety; General Operating Procedures; Emergency Contact Information; Part I; Manipulation of DNA; Lab Session 1; Getting Oriented; Practicing with Micropipettes; Station Checklist; Micropipetting; Laboratory Exercise; Preparing BSA Dilutions; Performing a Nitrocellulose Spot Test; Lab Session 2; Large-Scale Purification of Plasmid DNA; Introduction; Laboratory Exercises
Alkaline Lysis and Anion Exchange ChromatograhyDNA Quantification; Lab Session 3; Preparation of Expression Vector DNA (pET-41a(+), a GST Fusion Protein Vector); Introduction; Laboratory Exercises; Restriction Digestion of Vector (pET41a) Restriction Enzyme Digestions; Agarose Gel Electrophoresis; Cleaning DNA Using a Spin Column; Lab Session 4; Preparation of Insert DNA (egfp); Introduction; Laboratory Exercises; Restriction Digestion of pEGFP-N1; Isolation of egfp DNA from Agarose; Lab Session 5; Preparation of Transformation-Competent Cells and Control Transformation; Introduction
Laboratory Exercises Preparation of Chemically Competent Cells by Calcium Chloride Treatment; Transformation Control; Lab Session 6; DNA Ligation and Transformation of Escherichia coli; Introduction; Laboratory Exercises; Ligations and Ligation Controls; Divalent Cation-Mediated Transformation; Electrophoresis of Ligation Reactions; Part II; Screening Transformants; Lab Session 7; Colony Hybridizations; Lab Session 7A; Interim Laboratory Session; Introduction; Laboratory Exercises; Counting Transformants; Replica Plating; Lab Session 7B; Colony Hybridization: DNA Probe; Introduction
Laboratory Exercises Colony Hybridization with an egfp DNA Probe: Part 1; Labeling of DNA Probe by PCR Using Digoxigenin-11-dUTP; Lab Session 7C; Colony Hybridization: Monoclonal Antibody Probe; Introduction; Laboratory Exercise; Colony Hybridization with an a-GFP Monoclonal Antibody Probe: Part 1; Lab Session 8; Completion of Colony Hybridization with DNA Probe; Introduction; Laboratory Exercise; Colony Hybridization with an egfp DNA Probe: Part 2; Lab Session 9; Characterization of Recombinant Clones; Lab Session 9A; Completion of Colony Hybridization with mAB Probe; Introduction
Laboratory Exercise Colony Hybridization with an a-GFP monoclonal Antibody Probe: Part 2; Lab Session 9B; PCR Screening; Introduction; Polymerase Chain Reaction; Laboratory Exercise; Polymerase Chain Reaction Screen for Recombinant Clones: Part 1; Lab Session 9C; Visualization of Green Fluorescent Protein: Part 1; Introduction; Laboratory Exercise; Green Fluorescence Assay and Preparation of a Fresh Master Plate; Lab Session 10; Further Characterization of Recombinant Clones; Lab Session 10A:; Interim Laboratory Session; Laboratory Exercise; Inoculate Cultures for Minipreps; Lab Session 10B
Summary This manual is an indispensable tool for introducing advanced undergraduates and beginning graduate students to the techniques of recombinant DNA technology, or gene cloning and expression. The techniques used in basic research and biotechnology laboratories are covered in detail. Students gain hands-on experience from start to finish in subcloning a gene into an expression vector, through purification of the recombinant protein. The second edition has been completely re-written, with new laboratory exercises and all new illustrations and text, designed for a typical 15-week semester,
Bibliography Includes bibliographical references and index
Notes Print version record
Subject Recombinant DNA -- Laboratory manuals
Genetics -- Technique.
DNA, Recombinant -- analysis
DNA, Recombinant -- isolation & purification
Recombinant Proteins -- analysis
Recombinant Proteins -- isolation & purification
Genetic Techniques
SCIENCE -- Biotechnology.
Genetics -- Technique
Recombinant DNA
Genre/Form Laboratory manuals
Laboratory manuals.
Manuels de laboratoire.
Form Electronic book
Author Robertson, Dominique, 1951-
ISBN 0080456545