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•This course is introduced to bridge the gap between academics, research and industry. This course begins with a review of basic bio analytical technique and an introduction to general terminologies.
•This course contains bio analytical techniques along with their theory, working principal, common instrumentation and possible applications. This course will be equally beneficial to various scientific areas.
•Students will be exposed to various biological techniques and their applications in identification, isolation of different biological molecules.
• Students will know the principle and application of various instruments.
• Students will be able to make a strategy on molecular techniques for the improvement in any trait or its well being based on the techniques learned during this course.
• This course can use the knowledge for designing a project for research and execute it.
Module-1: Microscopic techniques
Visualization of cells and sub-cellular components by light microscopy and fluorescent microscope, Resolving powers of different microscopes, Electron microscope, Scanning and transmission microscopes, fixation and staining techniques for EM, Scanning probe microscopes: AFM and STM.
Practice: 1 To study and gain expertise on differential and cytological staining techniques.
Module-II: Spectroscopic techniques
Laws of absorption of light, Beer-Lambert’s Law, Absorption spectra, Measurement of absorption of light, Factors affecting the absorption properties of chromophores, Ultraviolet-visible absorption spectroscopy: Principle, Instrumentation and application, Fluorescence spectrophotometry: Principle, Instrumentation and application, Mass spectroscopy: Principle, Instrumentation and application.
Practice: 2 Demonstration of UV-vis Spectrophotometer.
Module-III: Radiolabeling Techniques
Isotopes and Nature of radioactivity, Radioactive decay, Radioisotopes used in Biology, Detection and measurement of radioactivity, Carbon dating, Geiger-Muller counting and liquid scintillation Counting, Safety guidelines related to Radiolabeling techniques.
Immunological techniques: Radio Immuno assay, ELISA, Double immune diffusion, Radial immune diffusion and immune precipitation.
Module-IV : Centrifugation techniques
Basic principles of sedimentation, Types of centrifuges, Types of rotors, Preparative centrifugation (Differential & density gradient), Analytical ultracentrifugation.
Practice:3 To separate proteins on the basis of their size and charge
Principles of chromatography (Adsorption and Partition chromatography), Planar chromatography (Paper and Thin-layer chromatography), Column chromatography,Gas chromatography, Gel permeation chromatography, Ion exchange chromatography,Affinity chromatography,HPLC
Practice:4 To separate the amino acids in a mixture by thin layer chromatography.
Practice:5 Purification of immunoglobulins by affinity chromatography
Module-VI: Electrophoretic techniques
General principles, Electrophoresis of nucleic acids (Agarose gel, pulse-field),Electrophoresis of proteins (SDS-PAGE, native gels)isoelectric focusing and two dimensional gels, Blotting techniques-Southern, northern,Western blotting.FISH and GISH
Practice:6 To study the separation of DNA by agarose gel electrophoresis
Module-VII: Electrophysiological Methods
Electrocardiogram (ECG), Positron emission tomography (PET), Nuclear Magnetic Resonance,Magnetic resonance imaging (MRI), Flow cytometry, Gene expression analysis. Sampling distribution; Regression and Correlation; t-test; Analysis of variance; Chi-square test.
Keith Wilson and John Walker (2009)Principles and techniques of biochemistry and molecular biology.7th Edition,Cambridge University Press, Cambridge, UK.
1. Keith Wilson and John Walker (2009)Principles and techniques of biochemistry and molecular biology.7th Edition,Cambridge University Press, Cambridge, UK.
2. Donald Voet and Judith Voet. Biochemistry, 4th Edition. (2010). John Wiley and Sons. New Jersey, USA
3. Rodney F Boyer(2012) Biochemistry laboratory: modern theory and techniques.2nd Edition, Pearson Prentice Hall, Boston,USA.
4. R. Katoch(2011) Analytical techniques in biochemistry and molecular biology,Springer, New York.
5. D. L. Spector, R. D. Goldman(2006), Basic methods in microscopy: protocols and concepts from cells: a laboratory manual.Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Radioisotopes in biologicalsystem
Ouchterlony Double Diffusion
Radial immuno diffusion
Sampling distribution; Regression and Correlation; t-test; Analysis of variance; Chi-square test.
Dr Sitaram Swain has received PhD degree from Utkal University in the year 2017. He is also trained and experienced from TATA Memorial Hospital, Mumbai, India. He has experienced with cell line culture, clinical biochemistry, immuno-fixation, immunoassay, cancer cytogenetic, serum protein electrophoresis and tumour marker analysis. He has qualified NET(ICAR-ASRB) in Animal biotechnology. He has […]