UID:
almahu_9948026250402882
Format:
1 online resource (303 p.)
ISBN:
1-281-31148-0
,
9786611311483
,
0-08-053683-2
Content:
This course manual instructs students in recombinant DNA techniques and other essential molecular biology techniques in the context of projects. The project approach inspires and captivates students; it involves them in the scientific experience, providing continuity to laboratory bench time and an understanding of the principles underlying the techniques presented. Molecular Biology is a must for any department, operating under budgetary constraints that offers or plans to offer a course in molecular cloning.Key Features* Includes a glossary of over 200 terms important
Note:
Description based upon print version of record.
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Front Cover; Molecular Biology: A Project Approach; Copyright Page; CONTENTS; METHODS LOCATOR; SUGGESTED SCHEDULE OF LABORATORY PROTOCOLS; PREFACE; ACKNOWLEDGMENTS; NOTE TO USERS; CHAPTER 1. TRANSPOSON MUTAGENESIS OF Escherichia coli; Introduction to Transposons; Advantages of Transposon Mutagenesis; Eukaryotic Transposable Elements; Transposons and Gene Fusions; Preparing for Laboratory Exercises; Tn5 Mutagenesis of Escherichia coil and Analysis of Auxotrophs: Overview; Protocol 1.1: Phage λ Titer; Protocol 1.2: Making a Phage Stock-Growing λ-Tn5'
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Protocol 1.3: Transposon Mutagenesis Using λ::TnphoA'-2Introduction to Auxotrophs; Protocol 1.4: Isolation of Auxotrophs-Replica Plating, Toothpicking, or Screening on 2 EM Plates; Protocol 1.5: Identification of Auxotrophs on Pool Plates; Protocol 1.6: Analysis of Auxotrophs Using a Literature Search; References; Suggested Reading; CHAPTER 2. RECOMBINANT DNA CLONING; Introduction to Recombinant DNA Technology; Cloning Vectors; Restriction Endonucleases; The Use of Restriction Endonucleases: Practical Matters; Ligase; Gel Electrophoresis; Ethidium Bromide Staining of DNA in Gels
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Ethidium Bromide SafetySensitivity of Detection with Ethidium Bromide; Other DNA Stains; Transformation; Recombinant DNA Cloning: Overview; Recombinant DNA: P1 Level of Physical Containment-Laboratory Practices; Protocol 2. la (Optional): Restriction Digestion of DNA Samples and Gel Electrophoresis of DNA Samples; Protocol 2.1 b (Optional): Restriction Enzyme Digestion of DNA to Be Cloned; Protocol 2.2 (Optional): Gel Electrophoresis; Protocol 2.3: Large-Scale Plasmid Isolation Using Alkaline Lysis; Protocol 2.4: Recombinant DNA Cloning; Protocol 2.5a: Competent Escherichia coli Cells
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Protocol 2.5b: Preparing Fresh Competent Escherichia coli Cells for TransformationProtocol 2.5c: A Rapid Colony Transformation Procedure; Protocol 2.6a: Boiling Mini-Prep Isolation of Plasmid DNA; Protocol 2.6b: Alkaline Mini-Prep Procedure for Isolating Plasmid DNA; References; Suggested Reading; CHAPTER 3. SOUTHERN BLOT ANALYSIS; Southern Blot Introduction; Using Southern Blot Analysis to Map Restriction Endonuclease Sites; Nonradioactive Labeling of Nucleic Acids; Autoradiography: Overview; Isolation of Nucleic Acid Fragments from Gels; Labeling Methods; Hybridization to Membranes
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The Attachment of Nucleic Acids to a MembraneProtocol 3.1 a: Southern Blot; Protocol 3.1 b: Bidirectional Blotting: A Sandwich Blot; Protocol 3.1 c: Alkaline Blotting; Protocol 3.1 d: Colony Hybridization; Protocol 3.2: Isolation of DNA Fragments by Electroelution; Protocol 3.3a: Labeling Probe with Biotin Using Nick Translation; Protocol 3.3b: Oligo Labeling of a Probe; Protocol 3.3c: Protobiotin Labeling of a Probe; Protocol 3.4a: Hybridization and Detection of Labeled Probe-A Biotin-Labeled Nonradioactive Probe and Chromogenic Substrate
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Protocol 3.4b: Hybridization and Detection of Labeled Probe-A Biotin-Labeled Nonradioactive Probe and Chemiluminogenic Substrate
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English
Additional Edition:
ISBN 0-12-397720-7
Language:
English
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