Biochemical Techniques
Teacher
Dr Sitaram Swain
Category
Core Courses
Course Attendees
Still no participant
Course Reviews
Still no reviews
Course Name : Biochemical Techniques
Code(Credit) : CUTM1697(3-2-1)
Course Objectives
- Students will learn about the principle and applications of spectrophotometry, different chromatographic techniques like gel filtration, Ion exchange, thin layer, etc.
Students will also learn about various electrophoretic techniques such as agarose, gel, PAGE, etc. and their applications in analyzing proteins and nucleic acids.
Students will learn the basic principles of centrifugation, various types of centrifuges, rotors and methods for subcellular fractionation. Principle and application of ELISA in biological experiments.
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.
Learning Outcomes
- The student will obtain hands-on training in basic separation techniques in biochemistry and gain expertise in the isolation of various biomolecules and organelles.
Students will know the principle and application of various instruments and they can be able to make strategy molecular techniques for the improvement in any trait or its well being based on the techniques learned during this course. Students can use this knowledge for biological experiments.
Course Syllabus
Module-I Chromatography: General Principles of chromatography, adsorption and partition. Techniques: Paper chromatography, Ascending, descending and circular, 2D chromatography, Rf values, column chromatography.
Module-II Principle of gel filtration, ion-exchange chromatography, affinity chromatography.
Thin layer chromatography (TLC) – Principle, procedure and applications.
Module-III Electrophoresis, Paper and gel electrophoresis, SDS-PAGE, 2D electrophoresis.
Module-IV Centrifugation: Principle of differential centrifugation. Types of Rotors, Ultra centrifuge –construction and applications in sub-cellular fractionation.
Module-V Radiochemistry: Natural and artificial radioactivity, characteristics of radioactive elements, units of radioactivity, disintegration constant, half-life, α, β and γ radiation. Detection of radioactivity by GM counter. Applications of radioisotopes – 3H, 14C, 131I, 60Co and 32P. Biological effects of radiations. Safety measure in handling radio isotopes.
Module-VI Types of ELISA, principle and application of ELISA,
Module-VII Spectrometry–Beer and Lamberts law, Principals of colorimeter, spectrophotometer. Nephelometry and turbidimetry and its applications
PRACTICAL CONTENT:
1. Identification of amino acids by paper chromatography.
2. Ascending paper chromatography of amino acids.
3. Separation of plant pigments using paper chromatography.
4. Separation of plant pigments by column chromatography using silica gel-G.
5. Polyacrylamide gel electrophoresis of proteins.
6. SDS-PAGE
7. Demonstration of separation of lipids by TLC.
8. Demonstration of two dimensional chromatography of amino acids
Recommended books:
Text Books:
Practical Biochemistry – Principles and Techniques, Wilson and Walker, Cambridge, University Press, Cambridge
Reference Books:
Biophysical Chemistry, Principles & Techniques – Upadhyay, Upadhyay and Nath Himalaya Publ. House.
Practical Biochemistry – Principles and Techniques, Wilson and Walker, Cambridge University Press, Cambridge
Boyer, R.F., Biochemistry Laboratory: Modern Theory and Techniques, 6th Ed., Boston, Mass: Prentice Hall, 2012.
Plummer D. T., An Introduction to Practical Biochemistry 3rd ed., Tata McGraw
Hill Education Pvt. Ltd. (New Delhi), 1998.
Session Plan
Session 1
Session 2
Adsorption and partition. Techniques: Paper chromatography
Session 3
Flipped class/Assignment:Adsorption and partition.Techniques: Paper chromatography
Session 10
Flipped Class/Assignment:Application of Principle of gel filtration chromatography
Session 11
Session 13
Ion-Exchange chromatography-animation
Principle, procedure and applications:Ion-exchange chromatography
Session 29
Types of Rotors, Ultra centrifuge –construction and applications in sub-cellular fractionation
Session 31
Radiochemistry: Natural and artificial radioactivity
Session 32
characteristics of radioactive elements, units of radioactivity, disintegration constant
Session 37
Assignment/ Flipped class-Applications of radioisotopes – 3H, 14C, 131I, 60Co and 32P
Session 38
Biological effects of radiations. Safety measure in handling radio isotopes.
Session 39
Flipped class-Biological effects of radiations and Safety measure in handling radio isotopes.
Session 50
Case Studies
Case Studies
Our Main Teachers
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 […]