|| BS21520 |
|| PRACTICAL SKILLS IN BIOCHEMISTRY |
|| 2001/2002 |
|| Dr David Hopper |
|| Semester 2 |
|| Dr David Lamb |
|| BS10910 - Introductory Biochemistry, BS12020 - Cell Structure, Metabolism & Function |
| Course delivery
|| Practical || 80 Hours (Up to twenty, 4-hour practical sessions) |
|| Seminars / Tutorials || 1 Hours |
|| Lab work and report || Continuous assessment of practical work throughout the module and presentation of selected experimental work in thesis format || 70% |
|| Exam || 2 Hours One 2-hour practical examination || 30% |
|| Resit assessment || 2 Hours One 2-hour practical examination (plus resubmission of failed or missing coursework or an alternative) || |
Aims and objectives
The aims of this module are to give students the opportunity to acquire the necessary skills to enable them to understand and interpret experimental protocols, and to carry out practical work to an acceptable level of accuracy with a variety of biological materials. This will be done by careful choice of experiments to ensure their exposure to a wide range of equipment and preparative practical procedures.
As a prelude to commencing experimental work students will attend an introductory session in which essential information about the course will be dealt with on an interactive basis. This will include aspects of laboratory safety, COSHH regulations, and the handling and presentation of experimental data.
The experimental work will be organised so that each student has the opportunity to experience a broad range of experimental procedures. Experiments offered are designed to familiarise the student with the laboratory facilities available and the range of equipment in routine use and, at the same time, develop qualitative and quantitative skills that will be essential for successful research project work at level 3.
Experiments can be grouped under the following general headings:
(1) Isolation of biological molecules. Purifications of the periplasmic enzyme, alkaline phosphatase from Escherichia coli and of cytochrome c from heart tissue make use of ion exchange and molecular exclusion chromatography. Isolation of trehalose from yeast and glycogen from rat liver both exploit differential solvent extraction and isolation of D-a-tocopherol from egg yolk provides experience of partition chromatography.
(2) Analytical techniques. The isolation of these biological materials leads logically to the use of a range of different techniques for analysis and determination of purity. These include various procedures leading to spectrophotometric measurements, polyacrylamide gel electrophoresis of proteins and TLC. Enzymes, their assay and characterisation play a fundamental role in many areas of biochemistry and the alkaline phosphatase is further exploited in kinetic studies that provide an intensive exercise in quantitative spectrophotometry.
Other experiments, designed to provide 'hands on' experience of specialised equipment include gas-liquid chromatography of fatty acids, radioactive monitoring of the products of an enzymic reaction, assessment of the metabolic versatility of bacteria and metabolic pathway selection by respirometry, fluorimetric analysis of protein-lizard binding sites and the use of endonucleases in DNA fingerprinting.
On completion of this module students should
be able to interpret written experimental procedures
have attained acceptable skill levels in quantitative measurements of parameters that are routinely encountered in practical biochemistry
have gained experience in handling prokaryotic and eukaryotic experimental material and the isolation and analysis of biological molecules
be familiar with a range of apparatus and many of the techniques commonly used in biochemical experimentation
be aware of hazards that are routinely encountered and of the necessary precautions that must be taken.
** Multiple Copies In Hugh Owen
Dawes, E.A.. (1980)
Quantitative problems in biochemistry. 6th. Livingstone.
Price, N. C. & Dwek, R.A.. (1989)
Principles and problems in physical chemistry for biochemists. 2nd. Oxford: Oxfrod University Press