CloseHelpPrint
Kies de Nederlandse taal
Course module: UCSCICHEL5
UCSCICHEL5
Metabolic Biochemistry Lab
Course info
Course codeUCSCICHEL5
EC2.5
Course goals
Experience life in a biochemical laboratory setting and get acquainted with various techniques involved, and with some elementary theory connected to metabolic biochemistry.
Content
This course is an introduction to laboratory techniques in biomedical research with a strong focus on metabolism. The goal is to clarify the biochemistry that underlies physiology and pathology.
Via several practical cases, the skills taught in each lab day are linked to clinical diagnoses. The theory of each experiment is discussed in short introductory lectures. Thereafter, each lab-experiment has to written up in a pre-constructed lab report before the real experiments start.
During this course the students are introduced to the following techniques: 
  • tissue homogenisation
  • differential centrifugation
  • membrane disruption techniques
  • isolation of mitochondria
  • molecular sorting with salt and alcohol
  • pH determinations
  • enzyme purification by ammonium sulphate precipitation
  • desalting with gelfiltration column chromatography
  • cellulose acetate electrophoresis
  • spectrofotometric assays for enzyme activity and protein content
  • calculation of specific activity of enzymes
  • enzymatic substrate determinations
  • glycogen digestion with amylase
  • study some biochemical characteristics of mitochondria
Format
Day 1: Focus on macromolecules. Protein and glycogen content in liver.
Day 2 and 3: Focus on enzymes. Isolation and partial purification of an enzyme from heart and skeletal muscle to diagnose tissue damage in blood of patients.
Day 4: Focus on enzymes continued. Purified enzymes will be used in the determination of metabolites in horse blood to decide which of two horses could be a good price winner. 
Day 5 morning: Focus on conformation of electrostatical charge on proteins (pH buffering properties) and diagnosis by protein electrophoresis.
Day 5 afternoon: Focus on  cell-membrane stability. Disruption of erythrocytes by osmotic swelling, detergents, freeze-thawing and ultra-sonic sound. Testing of the osmotic resistance of erythrocytes could give information about for example particular immuno diseases or hidden blood losses.
Day 6 and 7: Preparations for experiment day 8; writing lab report.
Day 8: Isolation of mitochondria from chicken-liver.
Day 9: Active site visit to the veterinary biochemistry and cell biology research laboratories (second floor Nieuw Gildenstein building)  division; finalizing lab report

Attendance
Due to the short duration and intensive nature of the lab course, 100% attendance is required.
CloseHelpPrint
Kies de Nederlandse taal