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The purpose of this laboratory manual is to introduce undergraduate students to techniques used in biochemistry and molecular biology laboratories and ensure that they master the lab skills necessary to be competitive in the job market. We present a collection of fifteen experiments that teach students sterile techniques, accurate pipetting, centrifuge usage, PCR, DNA purification, protein expression and purification, HPLC, enzyme kinetics, equilibrium binding assays and introduction to 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.

This course includes several chapters on the latest advancements in bioinformatics: how to access genome databank, perform sequence alignments, design primers, to predict secondary and tertiary structure and to use protein visualization tools.

This course material is based on the textbook written by Gerczei and Pattison under Creative Commons 4.0 license.

Course Curriculum

PREFACE
Preface FREE 00:20:00
1. Introducing the Bacterial Antibiotic Sensor Mini Project
What are Antibiotics? 00:40:00
What is Bacterial Antibiotic Resistance? FREE 00:45:00
How Do the Bacteria Detect Antibiotics In Its Environment? 00:20:00
How Does the ykkCD Sensor Exert Its Function? FREE 00:30:00
What Do We Do During the Mini Project? 00:30:00
2. Identifying Conserved Elements in the Toxin Sensor and Designing Mutants to Test Whether They are Important for Function
Mini Project Flowchart FREE 00:40:00
Why is Sequence Conservation Important for Macromolecule Function, and How Do We Determine This? 00:35:00
Review of Nucleic Acid Properties 00:30:00
Identifying Conserved Sequence Elements 00:30:00
3. Designing Primers for Site-Directed Mutagenesis
Basics of Site-directed Mutagenesis 00:50:00
Quickchange Site-Directed Mutagenesis 00:50:00
4. Performing Site-Directed Mutagenesis
Performing Site-Directed Mutagenesis 00:50:00
5. Purifying Mutant Toxin Sensor DNA from Bacterial Cells and Evaluating its Quality Using Agarose Gel Electrophoresis and UV Spectroscopy
Purifying Mutant Toxin Sensor DNA from Bacterial Cells and Evaluating its Quality Using Agarose Gel Electrophoresis and UV Spectroscopy FREE 00:50:00
6. Preparing DNA Template for Mutant RNA Sensor Synthesis Using a Restriction Endonuclease
Preparing DNA Template for Mutant RNA Sensor Synthesis Using a Restriction Endonuclease 00:50:00
7. Synthesizing the ykkCD Mutant Toxin Sensor RNA in vitro
Synthesizing the ykkCD Mutant Toxin Sensor RNA in vitro 01:30:00
8. Purifying the ykkCD Mutant Toxin Sensor RNA and Evaluating its Purity Using Denaturing PAGE and UV spectrometry
Purifying the ykkCD Mutant Toxin Sensor RNA and Evaluating its Purity 01:30:00
9. Evaluating the Ability of the ykkCD Toxin Sensor to Recognize the Antibiotic Tetracycline Using Fluorescent Quenching
Evaluating the Ability of the ykkCD Toxin Sensor 01:30:00
10. Evaluating Antibiotic Binding to Blood Serum Albumin Using Fluorescence Spectroscopy
Evaluating Antibiotic Binding to Blood Serum Albumin 01:30:00
11. Understanding the Importance of Buffers in Biological Systems
Importance of Buffers in Biological Systems 01:30:00
12. Molecular Visualization of an Enzyme, Acetylcholinesterase
Molecular Visualization of an Enzyme, Acetylcholinesterase 01:30:00
13. Determining the Efficiency of the Enzyme Acetylcholine Esterase Using Steady-State Kinetic Experiment
Determining the Efficiency of the Enzyme Acetylcholine Esterase 01:30:00
14. Separation of the Phosphatidylcholines Using Reverse Phase HPLC
Separation of the Phosphatidylcholines 01:30:00
TAKE THIS COURSE
  • FREE
  • 6 Weeks
4 STUDENTS ENROLLED
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