Molecular Biology Techniques - 4th Edition - ISBN: 9780128157749

Molecular Biology Techniques

4th Edition

A Classroom Laboratory Manual

Authors: Sue Carson Heather Miller Melissa Srougi D. Scott Witherow
Paperback ISBN: 9780128180242
Imprint: Academic Press
Published Date: 1st March 2019
Page Count: 275
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Description

Molecular Biology Techniques: A Classroom Laboratory Manual, Fourth Edition is a must-have collection of methods and procedures on how to create a single, continuous, comprehensive project that teaches students basic molecular techniques. It 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 will gain hands-on experience on subcloning a gene into an expression vector straight through to the purification of the recombinant protein.

Key Features

  • Presents student-tested labs proven successful in real classroom laboratories
  • Includes a test bank on a companion website for additional testing and practice
  • Provides exercises that simulate a cloning project that would be performed in a real research lab
  • Includes a prep-list appendix that 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

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 11A. Interim Laboratory Session
Lab Session 11B. Expression of Fusion Protein from Positive Clones, SDS-PAGE and Western Blot
Lab Session 12. Expression of Fusion Protein from Positive Clones, SDS-PAGE and Western Blot
Lab Session 13. Extraction of Recombinant Protein from Escherichia coli Using a Glutathione Affinity Column
Lab Session 13A. Interim Laboratory Session
Lab Session 13B. Extraction of Recombinant Protein from Escherichia coli and Purification Using a Glutathione Affinity Column
Lab Session 14. Analysis of Purification Fractions
Lab Session 14A. Analysis of Purification Fractions
Lab Session 14B. Replica Plate Positive Clone

Part 4. Analysis of mRNA Levels
Lab Session 15. Total RNA Purification
Lab Session 15A. Interim Laboratory Session
Lab Session 15B. Total RNA Purification
Lab Session 16. Analysis of gst::egfp mRNA Levels by RT-qPCR
Lab Session 17. Analysis of gst::egfp mRNA Levels by RT-qPCR
Lab Session 18. Analysis of gst::egfp mRNA Levels by Semi-Quantitative RT-PCR
Lab Session 19. Analysis of gst::egfp mRNA Levels by Semi-Quantitative RT-PCR

Appendix
1. Equipment
2. Prep List
3. Preparation of Competent E. coli Cells
4. Pre-Lab Questions

Details

No. of pages:
275
Language:
English
Copyright:
© Academic Press 2020
Published:
Imprint:
Academic Press
Paperback ISBN:
9780128180242

About the Author

Sue Carson

Affiliations and Expertise

North Carolina State University, Raleigh, U.S.A.

Heather Miller

Heather Miller

Dr. Heather Miller is an Assistant Professor of Biochemistry in the Chemistry Department at High Point University (High Point, NC). She graduated from Clarion University of Pennsylvania (Clarion, PA) with a B.S. in Molecular Biology/Biotechnology, and from Duke University (Durham, NC) with a Ph.D. in Molecular Genetics and Microbiology. She completed a teaching postdoctoral position in the Biotechnology Program at North Carolina State University (Raleigh, NC). Her area of scientific expertise is RNA biology. Her research focuses on HIV-1 gene expression and the coupling of transcription and splicing in humans. She has taught at the college level for nine years, and is engaged in the scholarship of teaching and learning.

Affiliations and Expertise

Assistant Professor of Biochemistry, Chemistry Department, North Carolina State University, Raleigh, USA

Melissa Srougi

Dr. Melissa C. Srougi is an Assistant Professor of Biochemistry at High Point University. She graduated from the University of Toledo (Toledo, OH) with a B.S. in Biology, and from Case Western Reserve University (Cleveland, OH) with a Ph.D. in Pharmacology. Her scientific areas of expertise are in experimental cancer chemotherapeutic agents and mechanisms of chemotherapeutic resistance. Dr. Srougi actively trains undergraduate research students in her laboratory. In addition, she has developed and taught a variety of inquiry-based college level science courses and has published and presented a number of peer-reviewed papers in the scholarship of teaching and learning.

Affiliations and Expertise

Assistant Professor of Biochemistry, High Point University

D. Scott Witherow

Dr. Witherow graduated from Rollins College (Winter Park, FL) with an A.B. in Chemistry, and from the University of Miami (Miami, FL) with a Ph.D. in Molecular and Cellular Pharmacology. His research has focused primarily on G protein-mediated signal transduction processes in mammalian systems. Following two research postdoctoral fellowships, Dr. Witherow served as a teaching postdoctoral fellow at North Carolina State University, where he published and presented multiple papers in the field of science education and developed a passion for teaching

Affiliations and Expertise

North Carolina State University, Raleigh, USA

Reviews

Review of a previous 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, 2012

Ratings and Reviews