Course

Unit 1: Spectroscopic and Biophysical Analysis of Biomolecules

Fluorescence Spectroscopy and FRET (Förster Resonance Energy Transfer) for biomolecular interactions, Circular Dichroism (CD) Spectroscopy for secondary structure analysis, NMR Spectroscopy for structure and dynamics of biomolecules, Mass Spectrometry in protein and nucleic acid analysis, Isothermal Titration Calorimetry (ITC) and Differential Scanning Calorimetry (DSC) for interaction studies

Unit 2: Advanced Chromatographic and Proteomics Tools

Affinity Chromatography and Fast Protein Liquid Chromatography (FPLC) for tagged protein purification High Performance Liquid Chromatography (HPLC) for biomolecule separation, 2D-Gel Electrophoresis and Mass Spectrometry-based Proteomics, Thermophoresis and Surface Plasmon Resonance (SPR) for kinetic and affinity profiling

Unit 3: Advanced Imaging and Cell Analysis Techniques

Confocal Laser Scanning Microscopy for 3D cellular imaging, Flow Cytometry and Fluorescence-Activated Cell Sorting (FACS), Fluorescence In Situ Hybridization (FISH) for nucleic acid localization, Super-Resolution Microscopy (STORM, PALM), Live Cell Imaging and Label-Free Imaging Techniques

Unit 4: Nucleic Acid Analysis and Genomic Tools

Digital PCR (dPCR) and Droplet Digital PCR (ddPCR) for ultra-sensitive nucleic acid quantification, FRET-based Nucleic Acid Hybridization Assays, Next-Generation Sequencing (NGS) and RNA-Seq, CRISPR-based diagnostics (e.g., SHERLOCK, DETECTR), ChIP-Seq, ATAC-Seq, and Bisulfite Sequencing for epigenomic profiling, Microarrays and Yeast Two-Hybrid for interaction mapping

LEARNING OBJECTIVES:

1. Understand the theoretical principles underlying advanced biophysical, spectroscopic, chromatographic, and genomic techniques used in modern biological research.
2. Learn to select and apply appropriate analytical tools for studying biomolecular structure, interactions, dynamics, and cellular localization.
3. Develop competence in interpreting experimental data from tools such as fluorescence spectroscopy, NMR, mass spectrometry, flow cytometry, and NGS.
4. Explore integrated applications of these techniques in proteomics, genomics, molecular diagnostics, and live-cell analysis.

COURSE OUTCOMES:

After completing the course, students shall be able to

CO1: Apply biophysical and spectroscopic methods to investigate biomolecular structure, folding, and interactions.

CO2: Demonstrate proficiency in protein purification, proteomics, and affinity/kinetic analysis using advanced chromatographic and biophysical tools.

CO3: Employ imaging and flow-based techniques for detailed cellular and molecular analysis, including real-time and high-resolution applications.

CO4: Analyze complex genomic and transcriptomic data using digital PCR, NGS, and CRISPR-based diagnostic tools to draw meaningful biological conclusions.

Reference

1.      Principles and Techniques of Biochemistry and Molecular Biology. Editors: Keith Wilson and John Walker
 Publisher: Cambridge University Press

2.      Introduction to Proteomics: Tools for the New Biology.Author: Daniel C. Liebler
 Publisher: Humana Press