UNIVERSITY OF ZIMBABWE

2017 Nov/Dec Examinations

Science
Faculty:...................................................................................................................................
Biochemistry
Department:............................................................................................................................
HBC201: Practical Techniques in Biochemistry 2
Paper code and Title:............................................................................................................
2 hours
Duration:.................................................................................................................................
Mr A. Kasiyamhuru
Examiner:................................................................................................................................

Authorized Materials: Calculator, Graph Paper

INSTRUCTIONS:

1. This paper contains 6 Questions

2. Answer 4 questions
3. Start each question on a new page
4. This question paper comprises 1 printed page

NB: DO NOT TURN OVER THE QUESTION PAPER OR COMMENCE WRITING UNTIL
INSTRUCTED TO DO SO.
1.
(a) Briefly describe the components of a flow cytometer and the principle of the method.
[20 marks]

(b) List the general uses of the flow cytometry technique. [5 marks]

2.
(a) State the composition of human blood and outline how lymphocytes can be obtained
from blood. [10 marks]

(b) Draw a labelled schematic diagram of a High Performance Liquid Chromatograph

(HPLC). [7 marks]

(c) Briefly describe the principles of the HPLC method of analysis. [8 marks]

3. With the aid of a labelled schematic diagram, describe the principles of Gas Liquid
Chromatography (GLC), including the applications and disadvantages of the method.
[25 marks]

4. Write an account of the principles of sodium dodecylsulphate – polyacrylamide gel

electrophoresis (SDS-PAGE), including the applications, advantages and disadvantages of
the technique. [25 marks]

5. Paying attention to the basis of separation of biomolecules using gel filtration and ion
exchange chromatography, discuss how these two chromatographic techniques can be used
in tandem to isolate a protein with a pI of 5 and a molecular weight 30 kDa.
[25 marks]
6. With reference to chromatographic techniques, distinguish between:

(a) mobile and stationary phase [4 marks]

(b) adsorptive and non-adsorptive technique [6 marks]
(c) gravity feed and pumping system [4 marks]
(d) total volume and void volume [5 marks]
(e) apparent and actual molecular weight. [6 marks]

END OF QUESTION PAPER