National Institute of Technology Rourkela

राष्ट्रीय प्रौद्योगिकी संस्थान राउरकेला

ଜାତୀୟ ପ୍ରଯୁକ୍ତି ପ୍ରତିଷ୍ଠାନ ରାଉରକେଲା

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Syllabus

Course Details

Subject {L-T-P / C} : LS3098 : Life Science Laboratory - VII { 0-0-3 / 3}

Subject Nature : Practical

Coordinator : Suman Jha

Syllabus

Module 1 :

The course contains a series of 8-10 experiments employing different analytical tools to characterize purified molecules. The design of the experiments is to give the students first-hand experience with expression/extraction and characterization of biomolecules/small bioactive molecules of interest.
Module I: Expression of recombinant human protein (?-Synuclein), and the analysis of the expression using UV-visible spectroscopy (for IPTG induction), PAGE (for expression of different proteins in the host).
Module II: Purification of the expressed protein using Ion Exchange Chromatography
Module III: Structural dynamics in purified protein (time/additive-based) using steady-state fluorescence spectroscope (including extrinsic/intrinsic fluorescence and fluorescence anisotropy).
Module IV: Structural dynamics in purified protein (time/additive-based) using Circular Dichroism Spectropolarimeter.
Module V: Thermodynamics of protein binding to different moieties/surfaces using Isothermal Titration/Differential Scanning Calorimeter(s).

Course Objective

1 .

The main objective, in brief, of the course is to give the students hands-on experience with different analytical tools employed by pharmaceutical/biotechnological industries and academic groups working in biological sciences. Hence, the specific objectives of the course are to:
1. Familiarize the students with the handling of analytical instruments and data analysis,

2 .

2. Understand different thermodynamic parameters’ role in governing different structures (molecular/population heterogeneity) to biomolecules,

3 .

3. Impart knowledge necessary to design and interpret the experiment's outcome using studied analytical tools.

Course Outcome

1 .

After completion of the course, the student will be able to:
? Categories different studied techniques as per their underlying principle and utilize them for designing the experiment/project,
? Tailor different specifications of the instrument as per the need of the project, and
? Understand the strengths and limitations of the techniques and use them in the design, execution, and interpretation of the outcomes from the experiment.

Essential Reading

1 .

Keith Wilson, John Walker, Practical Biochemistry (Principles and techniques),, Cambridge University Press,

2 .

Arrondo JLR, Alonso A (Eds.), Advanced Techniques in Biophysics, Springer (Series in Biophysics)

Supplementary Reading

1 .

J. A. Glasel, and M. P. Deutscher, Introduction to Biophysical Methods for Protein and Nucleic acid Research, Academic Press