Key Features

*Cover basic concepts and techniques used in molecular biology research labs *Student-tested labs proven successful in a real classroom laboratories *Exercises simulate a cloning project that would be performed in a real research lab *"Project" approach to experiments gives students an overview of the entire process *Prep-list appendix contains necessary recipes and catalog numbers, providing staff with detailed instructions

Readership

Graduate and undergraduate students studying biochemistry, molecular biology, biotechnology, and cell biology

Table of Contents

Preface

About the Authors

Acknowledgements

Note to Instructors

Instrumentation

Nomenclature

Introduction

Part 1. Manipulation of DNA

Lab Session 1. Getting Oriented

Lab Session 2. Purification and Digestion of Plasmid (Vector) DNA

Lab Session 3. PCR Amplification of egfp and Completion of Vector Preparation

Lab Session 4. Preparation of Insert DNA (egfp) PCR Product

Lab Session 5. DNA Ligation and Transformation of Escherichia coli

Part 2. Screening Transformants

Lab Session 6. Colony Hybridization

Lab Session 6A. Interim Laboratory Session

Lab Session 6B. Colony Hybridization: Monoclonal Antibody Probe

Lab Session 7. Characterization of Recombinant Clones

Lab Session 7A. Completion of Colony Hybridization with a Monoclonal Antibody Probe

Lab Session 7B. PCR Screening

Lab Session 7C. Prepare Fresh Replica Plate

Lab Session 8. Characterization of Recombinant Clones

Lab Session 8A. Interim Laboratory Session

Lab Session 8B. Analysis of PCR Screen Results

Lab Session 8C. Isolation of Miniprep DNA from Potential Transformants

Lab Session 8D. Visualization of Green Fluorescent Protein: Part 1

Lab Session 9. Characterization of Recombinant Clones

Lab Session 9A. Characterization of Miniprep DNA from Potential Transformants (Restriction Enzyme Analysis of Putative Transformants)

Lab Session 9B. Visualization of Green Fluorescent Protein: Part 2

Lab Session 9C. Computational Analysis of DNA Sequence from a Positive Clone: Part 1

Lab Session 10. Computational Analysis of DNA Sequence from a Positive Clone

Part 3. Expression, Detection and Purification of Recombinant Proteins from Bacteria

Lab Session 11. Expression of Fusion Protein from Positive Clones, SDS-PAGE and Western Blot

Lab Session

Details

No. of pages:
232
Language:
English
Copyright:
© 2012
Published:
Imprint:
Academic Press
Print ISBN:
9780123855442
Electronic ISBN:
9780123855459

About the authors

Heather Miller

Affiliations and Expertise

North Carolina State University, Raleigh, USA

D. Scott Witherow

Affiliations and Expertise

North Carolina State University, Raleigh, USA

Reviews

"Overall, this manual represents an invaluable training material on practical molecular biology for undergraduates, graduates, and inexperienced researchers. It could also introduce more experienced researchers to experiments that they have not considered previously."--Science Progress, vol 95, issue 2, 2012
"Whilst molecular biology has been the focus of course curricula in various bioscience educational programmes, there has been a lack of well-designed laboratory manuals to recommend for the practical sessions of these courses. The third edition of ‘Molecular Biology Techniques’ is one such excellent classroom laboratory manual. It encompasses experiments for 19 laboratory sessions presented as a semester-long project that gets students involved in a comprehensive experimental story from gene cloning to protein purification. The authors have employed the versatility of the PCR technique in various experiments and have also taken advantage of the enhanced green fluorescent protein in visualising positive clones. A new section involving five laboratory sessions on measuring mRNA levels has been added to this third edition. Overall, this manual represents an invaluable training material on practical molecular biology for undergraduates, graduates, and inexperienced researchers. It could also introduce more experienced researchers to experiments that they have not considered previously."--Science Progress