Syllabus

1. Visualization of protein structure by using PYMOL software.
2. Training on pH meter and preparation of Phosphate buffer of desired pH.
3. Determination of isoelectric point (pI) of amino acids (glycine and histidine) by titration method.
4. Theoretical estimation of isoelectric point and computing electrostatic surface of proteins by using PYMOL.
5. Amino acid analysis by Ninhydrin test.
6. Purification of protein by using ion exchange/size exclusion chromatography.
7. Estimation of the concentration of DNA and protein by spectrophotometric assay.
8. Determination of unknown concentration of protein using Bradford reagent (coomassie brilliant blue G250).
9. To estimate of concentration and size of DNA by agarose gel electrophoresis.
10. To differentiate D & L-Tryptophan and estimate secondary structures of protein by circular dichroism (CD).
11. Estimation of melting temperature of protein by far-UV circular dichroism.
12. Analysis of protein by steady state fluorescence spectroscopy.
13. Determination of mid-point potential (E1/2) of biological important analytes by cyclic voltammetry.
14. Monitoring enzymatic activity by oximeter based amperometry.
15. Detection and estimation of thiols by 4-DPS (4,4'-dithiodipyridine) or DTNB (5,5'-dithiobis-(2-nitrobenzoic)
Acid) assay.

To gain hands-on practice on biochemical techniques and bimolecular spectroscopies.

To highlights the concept of molecular/chemical biology, biochemical techniques and bimolecular spectroscopies.

1. At the end, student will have a real feeling about the biological macromolecules like, protein and DNA.
2. Students will be able to visualize, characterize and use these macromolecules (DNA and protein) for more complex biochemical assay.
3. Students will gain hands-on practice on different biochemical techniques and bimolecular spectroscopies.

Michael G. Gore, Spectrophotometry and Spectrofluorimetry: A Practical Approach, Oxford University Press

T.A. Brown, Essential Molecular Biology, Oxford University Press

Methods in Molecular Biology

Journal of Chemical Education