This workshop (6 hours/day) aims to introduce students (up to 8) to the required knowledge and practical hands-on skills necessary for preparatory and analytical research with proteins in a laboratory setting. Students will perform recombinant protein expression, protein purification and analysis of proteins with modern spectroscopic methods in biophysics. The theory and application of the following techniques will be covered: UV-visible absorbance spectroscopy, circular dichroism, fluorescence, single molecule, nuclear magnetic resonance and mass spectroscopy. At the conclusion of the module, students will have a good understanding of how these methods are used to study the structure of biomolecules. No prerequisites required.
The workshop provides a practical hands-on experience that covers some of the most important techniques applicable for experimental research in biophysics. CCBM fellows will be instructed on the preparatory and analytical skills necessary for working with proteins and develop a basic understanding of methods that are important in biophysical sciences. The course includes extensive hands-on laboratory work for two weeks.
The Summer Module is conjoined with BEST/Phys 231 graduate course part of the IB3 graduate emphasis organized by CCBM.
The general objectives of the workshop are:
1) Become familiar with the techniques that allow to purify, characterize and manipulate proteins.
2) Explore the fundamentals of spectroscopy and imaging techniques
Summary of the topics:
Recombinant Protein Expression and Purification
- Introduction to introduction to the structure and stability of biomolecules
- Growth and Expression of recombinant protein
- SDS-Page Electrophoresis
- Purification and analysis
Modern Spectroscopic Methods in Biophysics
- Introduction to spectroscopic techniques (UV/Vis spectroscopy).
- Circular Dichroism Spectroscopy
- Fluorescence Spectroscopy
- Single Molecule Spectroscopy
- Introduction to Nuclear Magnetic Resonance (NMR) spectroscopy
- Introduction to Mass Spectroscopy
Topics
- Protein Expression and Purification (Pre-inoculum + DNA transformation, bacterial growth)
- Protein Expression and Purification (Harvest the Cells, UV-Vis Spectroscopy + Buffer preparations)
- Protein Expression and Purification (Cell lysis and Electrophoresis)
- Protein Expression and Purification (Purification HPLC)
- Mass Spectroscopy
- Circular Dichroism Spectroscopy
- Fluorescence Spectroscopy (Steady state, FRET, TCSPC, Anisotropy)
- Single Molecule Experiment / TIRF
- NMR Spectroscopy
