P3: BIOCHEMICAL TECHNIQUES / SEM-III / SIDDY UNIT-I: CHROMATOGARPHIC METHODS. Principles & applications of techniques based on partitioning.

tioning. Counter current distribution. Paper chromatography (ascending, descending, two dimensional, circular). Thin layer chromatography- types of adsorbent, detection methods. Reversed phase chromatography. Gas chromatography. Methods of detection-flame ionization. ECD.

APPLICATIONS IN SEPERATION & PURIFICATION OF CHOS, A.AS, PEPTIDES, LIPIDS & NUCLEIC ACIDS. UNIT II: CHROMATOGRAPHIC METHODS II Ion- exchange chromatography- principle, types of exchangers & exchange capacity. Elution methods (stepwise, linear & isocratic elution). Applications of ion exchange chromatography. Gel exclusion chromatography, types of matrices & applications. Methodology & application of affinity chromatography. HPLC & reverse phase techniques. Chromatography of chiral compounds.

APPLICATIONS IN SEPERATION & PURIFICATION OF A.AS, PEPTIDES, PROTEINS, ENZYMES, NUCLEIC ACIDS, VITAMINS & PHARMACEUTICALS. UNIT III: SPECTROSCOPIC METHODS II Basic principles & applications of IR, ESR, NMR & Mass spectroscopy. Principles & applications of biophysical techniques such as ORD, CD & X-ray diffraction in relation to conformation analysis of bio molecules (outlines only). Instrumental aspects of design & operation.

UNIT IV: ELETROPHORETIC TECHNIQUES

Concept of electrophoresis- moving boundary & zonal electrophoresis, (paper ,cellulose acetate, gel). Principles & applications of native &SDS, PAGE, IEF, 2D. Capillary, High voltage, Agarose gel electrophoresis, Pulse field gel electrophoresis. Zymography. Blotting techniques- Southern, Northern & Western Blotting. APPLICATIONS IN SEPERATION AND ANALYSIS OF PEPTIDES, PROTEINS & NUCLEIC ACIDS