CLUSTER UNIVERSITY SRINAGAR
SYLLABUS (FYUP UNDER NEP 2020)
Offered By Department of Biotechnology
Semester 4th (Major Course)
Course Title: Biotechniques
Course Code : UGBTC22J401 Max. Marks 100
Credits: 4 (Theory: 3, Practical: 1) Theory External: 60; Min Marks: 24
Contact Hrs: 75 (Theory: 45, Practical: 30) Theory Internal (Continuous Assessment): 15 Marks, Min Marks: 06
Practical Experimental Basis= 15, Min. Marks: 06
Practical Experimental (Continuous assessment) = 10, Min. Marks: 04
Objectives
This course will introduce students to basic principles of various techniques and working of equipment’s used
across biological sciences for Sustainable Research and Analysis.
Learning Outcomes
▪ A student will be able to;
➢ Understand the basics of centrifugation, the principles and operation of differential and density gradient
centrifugation, and the precise applications of an ultracentrifuge.
➢ Understand the concepts and applications underlying various chromatography types.
➢ Have good understanding of various procedures, such as 2D gel electrophoresis, native polyacrylamide gel
electrophoresis, SDS PAGE, and agarose gel electrophoresis.
➢ Recognize the basic parts of a microscope while studying the concepts and real-world applications for
visualizings, identifying, and measuring cells and their constituent parts.
➢ Acquainted with the spectrophotometer's operation and learn about its many applications.
UNIT 1: 15 Hrs
Centrifugation: General principle of centrifugation, sedimentation coefficient, preparative and analytical
centrifugation, differential centrifugation & density-gradient centrifugation, ultracentrifugation and its
applications.
Chromatography: Principle, working and applications of thin-layer chromatography, ion-exchange
chromatography, gel filtration and affinity chromatography. HPLC.
UNIT 2: 15 Hrs
Electrophoresis: General principle and types; Principle, procedure and applications of native poly acrylamide gel
electrophoresis, sodium dodecyl sulphate polyacrylamide gel electrophoresis, isoelectric focusing, two-
dimensional gel electrophoresis and agarose gel electrophoresis, pulse field gel electrophoresis.
Blotting techniques: Southern, northern & western blotting; PCR: principle, methodology and applications.
UNIT 3 15 Hrs
Microscopy: Principle, working and applications of light microscopy - bright-field, dark-field, phase-contrast,
fluorescence & confocal microscopy, electron microscopy- TEM and SEM; Staining-principle and procedure of
simple staining, negative staining & differential staining.
Spectroscopy: Beer-Lambert’s law. Principle, working and applications of ultraviolet/visible light spectroscopy
(UV/Vis spectroscopy).
UNIT 4 (Practical/Lab Course) 30 Hrs
1. Operation and Principle of Centrifugation
2. Seperation of Amina Acids by paper/TLC
3. Extraction of DNA from different source available.
4. Extraction of total protein from bacteria/ animal source.
5. Protein separation by SDS-Polyacrylamide Gel Electrophoresis (PAGE)
6. Agarose gel electrophoresis.
7. Demonstration of PCR.
SUGGESTED READING:
1. Principles and Techniques of Biochemistry and Molecular Biology: Wilson, K. and Walker, J,
Cambridge University Press.
2. Physical Biochemistry - Applications to Biochemistry and Molecular Biology: Freifelder, D., W. H.
Freeman and Company.
3. Molecular Cloning - A Laboratory Manual: Sambrook, J. and Russell, D. W., Cold Spring Harbor
Laboratory Press
