7 Synthesizing the ykkCD Mutant Toxin Sensor RNA in vitro

Chapters in this book

1. Frontmatter I
2. Preface V
3. Contents IX
4. 1 Introducing the Bacterial Antibiotic Sensor Mini Project 1
5. 2 Identifying Conserved Elements in the Toxin Sensor and Designing Mutants to Test Whether They are Important for Function 9
6. 3 Designing Primers for Site-Directed Mutagenesis 27
7. 4 Performing Site-Directed Mutagenesis 38
8. 5 Purifying Mutant Toxin Sensor DNA from Bacterial Cells and Evaluating its Quality Using Agarose Gel Electrophoresis and UV Spectroscopy 51
9. 6 Preparing DNA Template for Mutant RNA Sensor Synthesis Using a Restriction Endonuclease 64
10. 7 Synthesizing the ykkCD Mutant Toxin Sensor RNA in vitro 81
11. 8 Purifying the ykkCD Mutant Toxin Sensor RNA and Evaluating its Purity Using Denaturing PAGE and UV spectrometry 89
12. 9 Evaluating the Ability of the ykkCD Toxin Sensor to Recognize the Antibiotic Tetracycline Using Fluorescent Quenching 97
13. 10 Evaluating Antibiotic Binding to Blood Serum Albumin Using Fluorescence Spectroscopy 113
14. 11 Understanding the Importance of Buffers in Biological Systems 124
15. 12 Molecular Visualization of an Enzyme, Acetylcholinesterase 132
16. 13 Determining the Efficiency of the Enzyme Acetylcholine Esterase Using Steady-State Kinetic Experiment 146
17. 14 Separation of the Phosphatidylcholines Using Reverse Phase HPLC 161
18. List of Figures 171