Basic Molecular Protocols in Neuroscience: Tips, Tricks, and Pitfalls - 1st Edition - ISBN: 9780128014615, 9780128015278

Basic Molecular Protocols in Neuroscience: Tips, Tricks, and Pitfalls

1st Edition

Authors: John Corthell
eBook ISBN: 9780128015278
Paperback ISBN: 9780128014615
Imprint: Academic Press
Published Date: 16th April 2014
Page Count: 124
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Description

Basic Neuroscience Protocols: Tips, Tricks, and Pitfalls contains explanatory sections that describe the techniques and what each technique really tells the researcher on a scientific level. These explanations describe relevant controls, troubleshooting, and reaction components for some of the most widely used neuroscience protocols that remain difficult for many neuroscientists to implement successfully. Having this additional information will help researchers ensure that their experiments work the first time, and will also minimize the time spent working on a technique only to discover that the problem was them, and not their materials.

Key Features

  • Describes techniques in very specific detail with step-by-step instructions, giving researchers in-depth understanding
  • Offers many details not present in other protocol books
  • Describes relevant controls for each technique and what those controls mean
  • Chapters include references (key articles, books, protocols) for additional study
  • Describes both the techniques and the habits necessary to get quality results, such as aseptic technique, aliquoting, and general laboratory rules

Readership

Graduate and undergraduate students in biological and biomedical sciences, post-doctoral students and fellows, as well as all researchers seeking a practical resource on neurobiological methods.

Table of Contents

  • Introduction
  • Chapter 1. General Notes
    • Aseptic Technique
    • Aliquoting
  • Chapter 2. DNA and RNA Extraction Protocols
    • Phenol:Chloroform (TRIzol™) Extraction Notes
    • Phenol:Chloroform Extraction Protocol
    • RNA Extraction
    • DNA Extraction
    • Protein Extraction
    • Kit Extraction of RNA Notes
    • Solution Recipes
  • Chapter 3. Agarose Gel Electrophoresis
    • Electrophoresis Notes
    • Agarose Gel Electrophoresis Protocol
    • Solution Recipes
  • Chapter 4. Reverse Transcription (RT) and Polymerase Chain Reaction (PCR)
    • Reverse Transcription Notes
    • Polymerase Chain Reaction (PCR) Notes
    • End-Point PCR
    • Primer Design
    • Reverse-Transcription PCR (RT-PCR)
    • Quantitative PCR (Real-Time PCR or qPCR)
    • Quantitation Methods
    • Running a Plate and Data Analysis
    • PCR Controls
    • PCR Troubleshooting
  • Chapter 5. Antibodies and Titrations
  • Chapter 6. Protein Extraction
    • Protein Extraction Notes
    • Total Cytosolic/Membrane Protein Extraction Protocol
    • Subcellular Fractionation Protocol (Adapted from Two Papers)8,9
    • Subcellular Fractionation Protocol (Detergents)
    • Solution Recipes
  • Chapter 7. Immunoblotting (Western Blot)
    • Immunoblotting Notes
    • SDS-PAGE and Immunoblot Protocols
    • Coomassie Blue Protocol
    • Fast Green/Ponceau S Protocol
    • Dot Blot Protocol17–19
    • Immunoblot Controls
    • Immunoblot Troubleshooting10,20
    • Solution Recipes
  • Chapter 8. Immunoprecipitation
    • Immunoprecipitation Notes
    • IP Protocol
    • IP Controls
    • IP Troubleshooting
    • Solution Recipes
  • Chapter 9. Perfusion and Immersion Fixation
    • Perfusion Fixation Notes
    • Immersion Fixation Protocol
    • Perfusion Fixation Protocol
    • Sample Storage
    • Solution Recipes
  • Chapter 10. Immunohistochemistry
    • Immunohistochemistry Notes
    • Antibody Titration Protocol (Ni-DAB)
    • Antibody Titration Protocol (Immunofluorescence)
    • IHC Protocol
    • Cresyl Violet Counterstain (Nissl Stain) Protocol27
    • IHC Controls
    • Solution Recipes
  • Chapter 11. In Situ Hybridization
    • In Situ Hybridization Notes
    • ISH Probe Design for Oligonucleotide and PCR Probes
    • ISH Protocol (Developed from Two Sources30,31)
    • ISH Controls23,33
    • Solution Recipes
  • References

Details

No. of pages:
124
Language:
English
Copyright:
© Academic Press 2014
Published:
Imprint:
Academic Press
eBook ISBN:
9780128015278
Paperback ISBN:
9780128014615

About the Author

John Corthell

Dr. Corthell received a PhD in Neuroscience from Florida State University, where he studied circadian rhythms using whole-cell patch-clamp electrophysiology, as well as numerous molecular biology techniques, in the olfactory system. He has more than 10 years of experience in laboratory work and techniques in the biological and chemical sciences, and more than 5 years teaching. The author has cultivated these techniques over the years through reading, asking questions, and trial and error. Each of these techniques has been tested, performed, and in some cases optimized by the author himself. He enjoys reading, writing, learning, children, and big dogs. When not working, he longs to go SCUBA diving.

Affiliations and Expertise

University of Michigan, Ann Arbor, MI, USA