Biochemical Analysis 1400-225ABCH

Topics of presentations: Isolation of protoplasts, nucleus, chloroplasts, mitochondria, Golgi membranes, microsomes, DNA, RNA, proteins, amino acids, carbohydrates, lipids. Methods of separation and purification of biomolecules: adsorption chromatography, column chromatography (CC), thin-layer chromatography (TLC), gas chromatography(GL), high performance liquid chromatography (HPLC), ion exchange chromatography, affinity chromatography, hydrophobic interaction chromatography, immobilized metal ion affinity chromatography, solid phase extraction, molecular filtration, electrophoresis, dialysis, electrodialysis. VIS/UV and IR spectrophotometry, NMR and mass spectrometry, radiometry, spinning.
Practices: 1. Determination of molecular weight of proteins by molecular filtration. 2. Electrophoretic analysis of proteins in polyacrylamide gel. 3. Potentiometric titration of amino acids. 4.Determination of buffer capacity.5. Determination of pK of uracil by spectrophotometric methods.6 Isolation of protoplasts from leaf mesophyll of marigold. 7. Incorporation of CH314COONa into oleanolic acid in protoplasts isolated from marigold leaves.8. Application of CH314COONa in labelling of sterols in marigold shoots. 9. Examination of phenylalanine ammonia lyase activity from barley hypocotyls. 10. Influence of functional groups on chromatographic properties of selected compounds. 11. Determination of the structure of flavonoids by spectral analysis.