Techniques in Biotechnology, Molecular Biology& Biochemistry

Welcome to "Techniques in Biotechnology & Molecular Biology," a comprehensive course designed to equip you with essential skills in Biotechnology, Molecular Biology and Biochemistry. Whether you're a student, researcher, or industry professional, this course will guide you through six key techniques crucial in the field of Biotechnology & Molecular Biology. Two-dimensional Gel Electrophoresis (2D-GE): Two-dimensional gel electrophoresis (2D-GE) is a powerful laboratory technique used to separate complex mixtures of proteins.

It combines two separation methods:Isoelectric Focusing (IEF): Proteins are first separated based on their isoelectric point (pI), which is the pH at which a protein has no net charge. SDS-PAGE: In the second dimension, proteins are further separated according to their molecular weight by applying an electric field through a polyacrylamide gel that contains sodium dodecyl sulfate (SDS). The result is a gel with a two-dimensional pattern of spots, where each spot represents a distinct protein.

This technique is widely used in proteomics to analyze protein expression, post-translational modifications, and identify biomarkers. Affinity Chromatography: Affinity chromatography is a technique used to separate and purify specific molecules from a mixture based on their selective binding to a ligand. In this method, a column is packed with a solid matrix (often agarose or sepharose) that is covalently attached to a specific ligand, such as an antibody, enzyme, or small molecule.

When a sample mixture is passed through the column, molecules that have a specific affinity for the ligand will bind to the column, while others are washed away. The bound molecules can then be eluted by changing conditions (e. g.