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BIO 504 TECHNIQUES IN BIOCHEMISTRY                                                                               (2L +2P) III

Objective
  To impart theoretical and practical knowledge about various biochemical techniques used in biochemistry for isolation and characterization of cellular components.

Theory

UNIT I
  rinciples of enzyme assays and analysis of biomolecules; Centrifugation techniques-theory, types and applications.

UNIT II
  Principles and applications of Spectrophotometry, Mass spectroscopy.

UNIT III
  Chromatographic techniques-TLC, ion - exchange, gel - filtration, affinity, GLC and HPLC.

UNIT IV
  Electrophoresis techniques – native and SDS-PAGE, Isoelectric focusing, capillary electrophoresis and agarose gel electrophoresis.

UNIT V
  Concept of radioactivity, tracer techniques and their applications in biology, Scintillation counters, autoradiography, radiation hazards and safety measures.

UNIT VI
  Polymerase Chain Reaction (PCR); Immunochemical techniques, MALDI-TOF.

UNIT VII
  Ethical issues and professional responsibilities in research.

Practicals
  pH and buffer preparation; Fractionation of cell organelles; Soluble protein estimation by Lowry’s Method; Isolation, purification and distribution of  polysomes; Gel Chromatography; polyacrylamide gel electrophoresis; Separation of proteins by iso-electric focusing. Isolation of plant DNA, Agarose gel electrophoresis; Estimation of fatty acids by gas-liquid chromatography, Radioisotope analysis by liquid scintillation counter, PCR.

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