Course Details
Subject {L-T-P / C} : CY3703 : Chemical Biology Laboratory { 0-0-3 / 2}
Subject Nature : Practical
Coordinator : Prof. Rabindra Kumar Behera
Syllabus
Preparation of Phosphate buffer of desired pH, Determination of the isoelectric point of amino acid –glycine and histidine by titration method. Visualization of protein structure by using PYMOL software, theoretical estimation of isoelectric point and computing electrostatic surface of proteins by using PYMOL. Estimation of the concentration of DNA and protein by spectrophotometric, and Bradford assay (coomassie brilliant blue G250). Purification of protein by using ion exchange chromatography, Estimatation of concentration and size of DNA by agarose gel electrophoresis, Distinguish DNA from protein by Bradford reagent, heat and acid, Differentiation of D & L-Tryptophan by circular dichroism (CD). Estimation of secondary structures of protein by CD, Estimation of melting temperature of protein by far-UV circular dichroism.
Course Objectives
- To highlights the concept of molecular biology, biochemical techniques and bimolecular spectroscopies.
Course Outcomes
1. At the end, student will have a real feeling about the biological macromolecules like, protein and DNA. <br />2. Students will be able to visualize, characterize and use these macromolecules (DNA and protein) for more complex biochemical assay.
Essential Reading
- Michael G. Gore, Spectrophotometry and Spectrofluorimetry: A Practical Approach, Oxford University Press
- T.A. Brown, Essential Molecular Biology, Oxford University Press
Supplementary Reading
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Journal and Conferences
- Journal of Chemical Education