Molecular Biology Techniques
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Molecular Biology Techniques

A Classroom Laboratory Manual

4th Edition - March 5, 2019

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  • Authors: Sue Carson, Heather Miller, Melissa Srougi, D. Scott Witherow
  • eBook ISBN: 9780128157756
  • Paperback ISBN: 9780128180242

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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 I: Manipulation of DNA
    Advanced Alternatives Within Part I
    1. Getting Oriented; Practicing With Micropipettes
    2. Purification and Digestion of Plasmid (Vector) DNA
    3. Completion of Vector Preparation and Polymerase Chain Reaction Amplification of egfp
    4. Preparation of Insert DNA (egfp) PCR Product
    5. DNA Ligation and Transformation of Escherichia coli

    Part II: Screening Transformants
    Advanced Alternatives Within Part II
    6. Screening of Transformants, Part I
    7. Screening of Transformants, Part II
    8. Analysis of DNA Sequence From a Positive Clone, Part II

    Part III: Expression, Detection, and Purification of Recombinant Proteins from Bacteria
    Advanced Alternatives Within Part III
    9. Expression of Fusion Protein from Positive Clones, SDS_PAGE and Western Blot: Part I
    10. Expression of Fusion Protein from Positive Clones, SDS_PAGE, and Western Blot: Part II
    11. Extraction of Recombinant Protein From Escherichia coli Using a Glutathione Affinity Column
    12. Analysis of Purification Fractions

    Part IV: Analysis of mRNA Levels
    13. Total RNA Purification
    14. Analysis of gst::egfp mRNA Levels by RT-qPCR: Part I
    15. Analysis of gst::egfp mRNA Levels by RT-qPCR: Part II
    16. Analysis of gst::egfp mRNA Levels by Semiquantitative RT-PCR: Part I
    17. Analysis of gst::egfp mRNA Levels by Semiquantitative RT-PCR: Part II

    Part V: Modulation of Gene Expression
    18. Culturing Mammalian Cells
    19. Transient Transfection of Mammalian Cells
    20. RNAi-Mediated Knockdown of EGFP: Part I
    21. RNAi-Mediated Knockdown of EGFP: Part II
    22. RNAi-Mediated Knockdown of EGFP: Part III
    23. CRISPR-Mediated Knockout of EGFP: Part I
    24. CRISPR-Mediated Knockout of EGFP: Part II
    25. Advanced CRISPR: Part I
    26. Advanced CRISPR: Part II
    27. Interim Laboratory Session
    28. Advanced CRISPR: Part IV
    29. Advanced CRISPR: Part V
    30. Advanced CRISPR: Part VI

    Appendix
    A: Equipment
    B: Prep List 
    C: Preparation of Competent Escherichia coli Cells 
    D: Pre-Lab Questions

Product details

  • No. of pages: 294
  • Language: English
  • Copyright: © Academic Press 2019
  • Published: March 5, 2019
  • Imprint: Academic Press
  • eBook ISBN: 9780128157756
  • Paperback ISBN: 9780128180242

About the Authors

Sue Carson

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

Department of Chemistry, High Point University, High Point, NC USA

Melissa Srougi

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

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. He is currently an Associate Professor at The University of Tampa, where he has been teaching undergraduate students since 2011.

Affiliations and Expertise

Department of Chemistry, Biochemistry, and Physics, The University of Tampa, Tampa, FL USA

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