Course code:
1422B		 Course name:
Methods in Biochemistry

Academic year:

2012/2013.

Attendance requirements:

(1001A + 1029B) / 1401B

ECTS:

10

Study level:

basic academic studies

Study program:

Biochemistry: 3. year, winter semester, compulsory course

Teachers:

Marija Đ. Gavrović Jankulović, Ph.D.
full professor, Faculty of Chemistry, Studentski trg 12-16, Beograd

Zoran M. Vujčić, Ph.D.
full professor, Faculty of Chemistry, Studentski trg 12-16, Beograd

Assistants:

Nikola L. Lončar, Ph.D.

Marica M. Grujić

Hours of instruction:

Weekly: three hours of lectures + seven hours of labwork (3+0+7)

Goals:

Students learn basic techniques: preparation of extracts, proteins and nucleic acids, the determination of their concentration. General chromatographic and FPLC techniques and electrophoresis. Independent processing of literature data.

Outcome:

Preparing students for independent problem solving in isolation of proteins, nucleic acids and small molecules, their structural determination and characterization. Training for the first steps in scientific research.

Teaching methods:

Lectures, experimental exercises, tutorial. Student powerpoint presentation topics.

Extracurricular activities:

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Coursebooks:

Main coursebooks:

  1. R. Reed, D. Holmes, J. Weyers, A. Jones: Practical skills in biomolecular sciences, 2nd edition, Pearson, Prentice Hall, Harlow, 2003.
  2. Zoran Vujčić: Experimental Biochemistry Practicum, Rantek, 2002.

Supplementary coursebooks:

  1. Robert Scopes: Protein purification, 2000.
  2. D. M. Bollag, D. M. Rozycki, S. J. Edelstein: Protein methods, John Wiley and Sons, 2002.

Additional material:

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  Course activities and grading method

Lectures:

0 points (3 hours a week)

Syllabus:

  1. Preparation and stabilization of protein crude extracts. Sterilization by microfiltration, chaotropes and kosmotropes, buffers.
  2. Biosensors, aptamers. The determination of molecular weight of proteins.
  3. Desalination and concentration of proteins.
  4. Protein purifications: Precipitation and Chromatographic Methods.
  5. Ion exchange and affinity chromatography.
  6. Ion exchange and affinity chromatography.
  7. EBA, HIC, IMAC.
  8. FPLC - fast protein liquid chromatography.
  9. Electrophoresis and isoelectric focusing and immobilized pH gradient (IPG).
  10. Protein staining techniques and zymogram detection. Peptide mapping. Chemical modification of proteins.
  11. Transfer and detection of proteins on membranes. Affinity electrophoresis. 2D electrophoresis.
  12. Preparative electrophoresis. Capillary Electrophoresis.
  13. Proteomics - fundamentals. Sequencing of proteins. MALDI TOF. BLAST.
  14. Isolation, quantification, and methods of recombinant DNA. PCR. Expression and purification of recombinant proteins.
  15. Sequencing NA.

Labwork:

10 points (7 hours a week)

Syllabus:

1-2. Preparation of protein crude extracts and determination of protein concentration.

3-4. Protein precipitation.

5-6. Molecular weight determination (HRPO).

7-10. Ion exchange, HIC, IMAC, FPLC: superdex, HIC, Mono Q. - fast protein liquid chromatography.

10-12. Electrophoresis: native and isoelectric focusing SDS PAGE, UT electrophoresis. 2D electrophoresis.

13-14. Isolation, quantification of DNA. Agarose electrophoresis.

14-15. RNA, purification quantification, analysis, electrophoresis.

Written exam:

30 points

Oral exam:

40 points

Tutorial:

20 points