Integrated Laboratory Practicals III

Objectives

• Observe safety rules and regulations in the laboratory.
• Handle properly standard laboratory equipment.
• Apply safe laboratory rules and follow molecular biology and tissue culture protocols.
• Produce an ordered and intelligible record of the activities performed in the laboratory
• Design and use an experimental protocol.

Learning outcomes

1. Demonstrate skill in transfecting cells and evaluating transfection efficiency.
2. Recognise cell differentiation markers by western blot and know of other methodologies (RT-PCR, flow cytometry, etc.).
3.  Produce a monolayer cell culture using an established cell line.
4. Apply the basic techniques commonly used in molecular genetics such as cloning, transfection and transformation of a microorganism.
5. Use biochemical techniques for the evaluation of proteins, electrophoresis and immunodetection.
6. Analyse and interpret the data obtained.
7. Demonstrate comprehension of spoken and written messages of different types in Catalan, Spanish and English.
8. Prepare written reports and documents (mainly of a technical nature) with correct spelling and grammar in English.
9. Perform well in work situations with commitment and responsibility.

Competencies

General skills

• Combine scientific knowledge with technical skills and technological resources to deal with problems in professional practice.
• Endeavour to combine independence and personal initiative with teamwork in multidisciplinary activities.

Specific skills

• Have oral and written skills in English for communicating results, conclusions and processes deriving from biotechnology research and process management.
• Interpret results obtained in biotechnology laboratories on the basis of correct application of laboratory protocols and basic techniques, making appropriate use of suitable instruments, with due regard for established safety standards.
• Use the main techniques and methods for manipulation and modification of biological systems.

Basic skills

• Students have developed the learning skills necessary to undertake further studies with a high degree of independent learning.
• Students have the ability to gather and interpret relevant data (usually within their field of study) in order to make judgements that include reflection on relevant social, scientific and ethical issues.

Core skills

• Become the protagonist of one's own learning process in order to achieve personal and professional growth and acquire all-round training for living and learning in a context of respect for linguistic, social, cultural, gender and economic diversity.
• Use oral, written and audiovisual forms of communication, in one's own language and in foreign languages, with a high standard of use, form and content.

Content

• Permanent and transient transfection of cell lines
  • Fluorescence microscopy follow up
  • Drug resistance
• Cell differentiation. Culture of myoblasts in different media to achieve their proliferation or differentiation; signalling pathways involved
  • Differentiation marker analysis using western blot and enzyme activity tests
  • Morphology assessment under the microscope using specific staining

Evaluation

• Assessment activities that represent 100% of the final grade (NF):
  • Activity 1. Final written test (40%). Minimum pass mark 4/10 (may be retaken)
  • Activity 2. Group work oral .presentation (15%)
  • Activity 3. Poster with data obtained (15%). (Late submission penalisation: 20%)
  • Activity 4. Attitude in the lab and questionnaires (15%)
  • Activity 5. Final report (15%)
• Attendance to all sessions is compulsory.

Methodology

• This subject is developed through several guided laboratory sessions. Two demonstrative experiments are performed in groups of two or three people using different methodologies. Students are given a series of lab protocols that they must follow to achieve partial goals in the development of the project. The generation of reagents by the student is necessary to proceed from one step to another.
• There are also group discussions during the development of the experiments and discussions on two research articles related to the methodology and contents of the experiments performed. Students are required to prepare different parts of the articles and present them to the rest of the class.

Bibliography

Key references

• Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J.G., Smith, J. A., Struhl, K. (eds.) (2002). Short protocols in molecular biology (5 ed.). John Wiley & Sons.
• Blitterswijk,C.A. & Boer J. (2023). Tissue engineering. Retrieved from https://ucercatot.uvic-ucc.cat/permalink/34CSUC_UVIC/1nl2ep/alma991001143979106718

Further reading

Teachers will provide complementary bibliography and compulsory reading throughout the course via the Virtual Campus.