Biochemistry Laboratory Manual For Undergraduates: An Inquiry-Based Approach

Timea Gerczei Fernandez; Scott Pattison

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Biochemistry Laboratory Manual For Undergraduates

An Inquiry-Based Approach

Timea Gerczei Fernandez and Scott Pattison

Language: English

Published/Copyright: 2015

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About this book

Biochemistry laboratory manual for undergraduates – an inquiry based approach by Gerczei and Pattison is the first textbook on the market that uses a highly relevant model, antibiotic resistance, to teach seminal topics of biochemistry and molecular biology while incorporating the blossoming field of bioinformatics. The novelty of this manual is the incorporation of a student-driven real real-life research project into the undergraduate curriculum. Since students test their own mutant design, even the most experienced students remain engaged with the process, while the less experienced ones get their first taste of biochemistry research. Inclusion of a research project does not entail a limitation: this manual includes all classic biochemistry techniques such as HPLC or enzyme kinetics and is complete with numerous problem sets relating to each topic.

Biochemistry, Laboratory Manual, Molecular Biology

Author / Editor information

Timea Gerczei Fernandez, Ball State University, Muncie, USA
Scott Patison, Ball State University, Muncie, USA

Topics

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Frontmatter
I

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Preface
V

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Contents
IX

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1 Introducing the Bacterial Antibiotic Sensor Mini Project
1

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2 Identifying Conserved Elements in the Toxin Sensor and Designing Mutants to Test Whether They are Important for Function
9

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3 Designing Primers for Site-Directed Mutagenesis
27

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4 Performing Site-Directed Mutagenesis
38

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5 Purifying Mutant Toxin Sensor DNA from Bacterial Cells and Evaluating its Quality Using Agarose Gel Electrophoresis and UV Spectroscopy
51

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6 Preparing DNA Template for Mutant RNA Sensor Synthesis Using a Restriction Endonuclease
64

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7 Synthesizing the ykkCD Mutant Toxin Sensor RNA in vitro
81

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8 Purifying the ykkCD Mutant Toxin Sensor RNA and Evaluating its Purity Using Denaturing PAGE and UV spectrometry
89

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9 Evaluating the Ability of the ykkCD Toxin Sensor to Recognize the Antibiotic Tetracycline Using Fluorescent Quenching
97

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10 Evaluating Antibiotic Binding to Blood Serum Albumin Using Fluorescence Spectroscopy
113

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11 Understanding the Importance of Buffers in Biological Systems
124

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12 Molecular Visualization of an Enzyme, Acetylcholinesterase
132

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13 Determining the Efficiency of the Enzyme Acetylcholine Esterase Using Steady-State Kinetic Experiment
146

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14 Separation of the Phosphatidylcholines Using Reverse Phase HPLC
161

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List of Figures
171

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Publishing information

Pages and Images/Illustrations in book

eBook published on:

December 15, 2014

eBook ISBN:

9783110411331

Hardcover published on:

December 15, 2014

Hardcover ISBN:

9783110411324

Pages and Images/Illustrations in book

Main content:

186

Illustrations:

59

Coloured Illustrations:

19

Tables:

7

https://doi.org/10.2478/9783110411331

eBook ISBN: 9783110411331

Hardcover ISBN: 9783110411324

Audience(s) for this book

Undergraduate students and academic teachers

Creative Commons

BY-NC-ND 3.0

Safety & product resources

Manufacturer information:
Walter de Gruyter GmbH
Genthiner Straße 13
10785 Berlin
productsafety@degruyterbrill.com

Biochemistry Laboratory Manual For Undergraduates

Overview

About this book

Author / Editor information

Topics

Table of contents

Frontmatter

Preface

Contents

1 Introducing the Bacterial Antibiotic Sensor Mini Project

2 Identifying Conserved Elements in the Toxin Sensor and Designing Mutants to Test Whether They are Important for Function

3 Designing Primers for Site-Directed Mutagenesis

4 Performing Site-Directed Mutagenesis

5 Purifying Mutant Toxin Sensor DNA from Bacterial Cells and Evaluating its Quality Using Agarose Gel Electrophoresis and UV Spectroscopy

6 Preparing DNA Template for Mutant RNA Sensor Synthesis Using a Restriction Endonuclease

7 Synthesizing the ykkCD Mutant Toxin Sensor RNA in vitro

8 Purifying the ykkCD Mutant Toxin Sensor RNA and Evaluating its Purity Using Denaturing PAGE and UV spectrometry

9 Evaluating the Ability of the ykkCD Toxin Sensor to Recognize the Antibiotic Tetracycline Using Fluorescent Quenching

10 Evaluating Antibiotic Binding to Blood Serum Albumin Using Fluorescence Spectroscopy

11 Understanding the Importance of Buffers in Biological Systems

12 Molecular Visualization of an Enzyme, Acetylcholinesterase

13 Determining the Efficiency of the Enzyme Acetylcholine Esterase Using Steady-State Kinetic Experiment

14 Separation of the Phosphatidylcholines Using Reverse Phase HPLC

List of Figures

Bibliographic data