BT3011 Modern Bio Analytical Techniques Syllabus:

BT3011 Modern Bio Analytical Techniques Syllabus – Anna University Regulation 2021

OBJECTIVES:

 To study the various analytical techniques used in Biotechnology.

UNIT I SPECTROSCOPY STUDY OF CHEMICAL COMPOUNDS AND BIO-MOLECULES

Electromagnetic radiations and interactions with matters: Electromagnetic spectrum. Quantisation of energy, Electronic, vibrational and rotational spectroscopy. Franck–Condon principle, Jablonski diagram, radiative, nonradiative pathways, fluorescence and phosphorescence. Absorption of radiation, BeerLambert’s law, deviation of Beer-Lambert’s equation and its limitations. Principals, instrumentation, sampling and application of few spectroscopic techniques: UV-Visible spectroscopy, Fluorescence spectroscopy, IR/Raman spectroscopy, NMR Spectroscopy and Mass spectroscopy.

UNIT II DIFFRACTION TECHNIQUE

Introduction to lattice and lattice systems, Bragg’s plane, miller indices, point groups and space groups Principle of diffraction and X-ray diffraction: X-rays production, X- ray spectra, Bragg’s law and intensity of X- rays, Mosley’s law, powdered XRD, percentage crystallinity, single crystal XRD, macromolecular XRD (protein crystallization, data collection and structure solution).

UNIT III CHROMATOGRAPHY

Classification of chromatographic techniques and their principles, Theory of chromatography, band broadening, rate and plate theory factors responsible for separation. Column chromatography, TLC, Paper chromatography. Liquid Chromatography and HPLC: Instrumentation, pumps, solvent delivery system, isocratic and gradient programming modes, sample introduction system, columns, detectors, reversed phase and normal phase chromatography. Gas Chromatography: Instrumentation, carrier gas supply, injectors, columns, packed and capillary columns, column oven and temperature programming, different detectors. Introduction to hyphenated techniques in chromatography, GC-MS and LC-MS.

UNIT IV MICROSCOPY

Microscopy with light and electrons – Electrons and their interaction with the specimen – Electron, diffraction – Instrument, specimen preparation and application of TEM and SEM – Fluorescence microscopy – Laser confocal microscopy – Phase contrast – Video microscopy – Scanning probe microscopy.

UNIT V ELECTROPHORETIC TECHNIQUES

Principle, equipment and process, Agarose gel electrophoresis, horizontal and vertical gel electrophoresis, electrophoresis techniques, Isoelectric focusing, capillary electrophoresis and application of electrophoresis in analysing macromolecules.

TOTAL 45 PERIODS

OUTCOMES:

 The students will be capable of handling different instruments in the laboratory.
 They would be able to compare different separation techniques and use them effectively in research work

TEXTBOOKS:

1. D. Campbell, Biological spectroscopy (Benjamin/Cummings Pub. Co, Menlo Park, Calif, 1984), Biophysical techniques series. 2. K. Wilson, J. M. Walker, Eds., Principles and techniques of biochemistry and molecular biology (Cambridge University Press, Cambridge, UK : New York, 7th ed., 2009)
2. R. F. Boyer, Biochemistry laboratory: modern theory and techniques (Prentice Hall, Boston, 2nd ed., 2012).
3. R. Katoch, Analytical techniques in biochemistry and molecular biology (Springer, New York, 2011).
4. D. L. Spector, R. D. Goldman, Eds., Basic methods in microscopy: protocols and concepts from cells: a laboratory manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y, 2006).
5. R. L. Switzer, Experimental biochemistry (W. H. Freeman and Co, New York, 3rd ed., 1999).
6. Chandler, D. and Roberso, R.W., “Bioimaging: Current Techniques in Light & Electron Microscopy”, Jones and Bartlett publishers, 2008.

REFERENCE:

1. R. F. Boyer, Modern experimental biochemistry (Benjamin Cummings, San Francisco, 3rd ed., 2000).
2. J. R. Lakowicz, Principles of fluorescence spectroscopy (Springer, New York, 2006;
3. B. Fultz, Transmission electron microscopy and diffractometry of materials (Springer, Berlin ; New York, 2nd ed., 2002).
4. D. B. Williams, C. B. Carter, Transmission electron microscopy a textbook for materials science (Springer, New York, 2009
5. R. M. Silverstein, Spectrometric identification of organic compounds (John Wiley & Sons, Hoboken, NJ, 7th ed., 2005).
6. D. Harvey, Modern analytical chemistry (McGraw-Hill, Boston, 2000).
7. Pavia, D.L., Lampman, G.M., Kriz, G.S. and Vyvyan, J.R., “Introduction to Spectroscopy”, 4th Edition, Brooks/Cole Cengage Learning, 2008.