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Elsevier

Molecular Biology Techniques

A Classroom Laboratory Manual

  • 4th Edition - March 5, 2019
  • Newer edition is available
  • Authors: Sue Carson, Heather B. Miller, Melissa C. Srougi, D. Scott Witherow
  • Language: English

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, comprehen… Read more

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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 DNAAdvanced Alternatives Within Part I 1. Getting Oriented; Practicing With Micropipettes 2. Purification and Digestion of Plasmid (Vector) DNA3. Completion of Vector Preparation and Polymerase Chain Reaction Amplification of egfp 4. Preparation of Insert DNA (egfp) PCR Product5. DNA Ligation and Transformation of Escherichia coli

Part II: Screening TransformantsAdvanced Alternatives Within Part II6. Screening of Transformants, Part I 7. Screening of Transformants, Part II8. Analysis of DNA Sequence From a Positive Clone, Part II

Part III: Expression, Detection, and Purification of Recombinant Proteins from BacteriaAdvanced 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 Column12. Analysis of Purification Fractions

Part IV: Analysis of mRNA Levels13. Total RNA Purification14. Analysis of gst::egfp mRNA Levels by RT-qPCR: Part I15. 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 Expression18. Culturing Mammalian Cells19. Transient Transfection of Mammalian Cells20. RNAi-Mediated Knockdown of EGFP: Part I 21. RNAi-Mediated Knockdown of EGFP: Part II22. RNAi-Mediated Knockdown of EGFP: Part III23. CRISPR-Mediated Knockout of EGFP: Part I24. CRISPR-Mediated Knockout of EGFP: Part II 25. Advanced CRISPR: Part I 26. Advanced CRISPR: Part II27. Interim Laboratory Session 28. Advanced CRISPR: Part IV 29. Advanced CRISPR: Part V30. Advanced CRISPR: Part VI

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

Review quotes

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

Product details

About the authors

SC

Sue Carson

Dr. Susan Carson is a Professor of Plant and Microbial Biology at North Carolina State University (Raleigh, NC) where she has served on the faculty since 2001. She directs the Master of Microbial Biotechnology Program and also leads a university-wide faculty development program focused on enhancing students’ critical and creative thinking skills across disciplines. Prior to her current role, Dr. Carson spent over a decade leading curriculum development for the North Carolina State Biotechnology Program and two years as a Program Officer at the National Science Foundation Division of Undergraduate Education. Her current work focuses on college-level biology education, enhancing students’ higher order thinking across disciplines, and integration of Design Thinking in a Professional Science Masters (PSM) program. She graduated from Rutgers University (New Brunswick, NJ) with a BS in Biotechnology, and from the University of North Carolina (Chapel Hill, NC) with a PhD in Microbiology.

Affiliations and expertise
Department of Plant and Microbial Biology, North Carolina State University, Raleigh, USA

HM

Heather B. Miller

Dr. Heather B. Miller is an Associate Professor of Biochemistry at High Point University (High Point, NC). Her area of scientific expertise is RNA biology. Her research focuses on MRSA and other pathogens’ gene expression when challenged with antibiotics and novel antibiotic adjuvants. She has mentored over 30 undergraduate students and is the Principal Investigator of a National Institutes of Health (NIH) Academic Research Enhancement Award (AREA). She was named a Henry Dreyfus Teacher-Scholar for her outstanding accomplishments in research and education in the chemical sciences. She has developed and taught multiple biochemistry and biotechnology courses and has published and presented a number of peer-reviewed papers in the scholarship of teaching and learning. Dr. Miller graduated from Clarion University of Pennsylvania (Clarion, PA) with a BS in Molecular Biology/Biotechnology, and from Duke University (Durham, NC) with a PhD in Molecular Genetics and Microbiology. She completed a teaching postdoctoral position in the Biotechnology Program at North Carolina State University
Affiliations and expertise
Department of Chemistry, High Point University, High Point, NC, USA

MS

Melissa C. Srougi

Dr. Melissa C. Srougi is an Associate Professor in the Department of Molecular Biomedical Sciences and the Biotechnology Program at North Carolina State University (Raleigh, NC) with over 16 years of research experience in higher education. Her scientific areas of expertise are in experimental cancer chemotherapeutics as well as the scholarship of teaching and learning. She is passionate about expanding opportunities for undergraduate students to participate in research and actively trains undergraduate students in her laboratory. She is co-Principal Investigator of a National Science Foundation (NSF) funded Research Experience for Undergraduates Program and the Director of the Graduate Certificate in Molecular Biotechnology. Dr. Srougi has received funding from various federal and private agencies, including the NSF, BioMade, Department of Defense, Department of Education and the American Cancer Society. She teaches a wide variety of undergraduate/graduate interdisciplinary STEM lecture and lab courses focusing on biochemistry, cancer drug discovery, and biotechnology. Dr. Srougi serves as an Associate Editor for Frontiers in STEM Education. She has published numerous peer-reviewed papers on the scholarship of teaching and learning with a focus on collaborative learning, CUREs and metacognition. Dr. Srougi graduated from the University of Toledo (Toledo, OH) with a BS in Biology and from Case Western Reserve University (Cleveland, OH) with a PhD in Pharmacology.
Affiliations and expertise
Department of Molecular Biomedical Sciences and the interdisciplinary Biotechnology Program at NC State University

DW

D. Scott Witherow

Dr. D. Scott Witherow is an Associate Professor of Biochemistry at The University of Tampa (Tampa, FL). His research focuses primarily on the regulation of G-protein-mediated signal transduction processes and biochemical education and pedagogy. In addition to mentoring over 20 undergraduate research students, Dr. Witherow has designed and published biochemistry lab courses and has been recognized by the American Society for Biochemistry and Molecular Biology as an education fellow for his efforts in the areas of instruction and assessment. He graduated from Rollins College (Winter Park, FL) with an AB in Chemistry and from the University of Miami (Miami, FL) with a PhD in Molecular and Cellular Pharmacology. Following traditional postdoctoral research positions at Duke University, he transitioned into pedagogy through a teaching postdoctoral position at North Carolina State University
Affiliations and expertise
Department of Chemistry and Biochemistry, The University of Tampa, Tampa, FL USA

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