ean 9780123855442 temática BIOLOGÍA año Publicación 2011 idioma INGLÉS editorial ELSEVIER formato RÚSTICA 73,80 €    PEDIR   NOVEDAD

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CONTENTS Foreword Acknowledgements Note to Instructors Instrumentation Nomenclature INTRODUCTION: Conceptual outline for experiments PART I MANIPULATION OF DNA Lab session 1: Getting oriented; Practicing with pipetmen Lab session 2: Large scale purification of plasmid DNA Lab session 3: Preparation of expression vector DNA (pET-41a(+), a GST fusion protein vector) Lab session 4: Preparation of insert DNA (egfp) Lab session 5: Preparation of transformation-competent cells and control transformation Lab session 6: DNA ligation and transformation of Escherichia coli PART II SCREENING TRANSFORMANTS Lab session 7: Colony hybridizations Lab session 7a: Interim laboratory session: Lab session 7b: Colony hybridization: DNA probe Lab session 7c: Colony hybridization: Monoclonal antibody probe Lab session 8: Completion of colony hybridization with DNA probe Lab session 9: Characterization of recombinant clones Lab session 9a: Completion of colony hybridization with mAB probe Lab session 9b: PCR screen Lab session 9c: Visualization of green fluorescent protein: Part 1 Lab session 10: Further characterization of recombinant clones Lab session 10a: Interim laboratory session: Lab session 10b: Analysis of PCR screen results Lab session 10c: Isolation and characterization of miniprep DNA from potential transformants (Restriction analysis of putative transformants) Lab session 10d: Visualization of green fluorescent protein: Part 2 PART III EXPRESSION, DETECTION, AND PURIFICATION OF RECOMBINANT PROTEINS FROM BACTERIA Lab session 11: Expression of fusion protein from positive clones and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Immunological analysis (Western blot): Part 1 Lab session 12: Expression of fusion protein from positive clones and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Immunological analysis (Western blot): Part 2 Lab session 13: Extraction of recombinant protein from Escherichia coli using a glutathione affinity column Interim laboratory session: I. Laboratory exercise: Inoculate cultures for protein purification Lab session 14: Analysis of purification fractions Appendices: Equipment, Prep list, Making sense of orientation