Practical classes of Molecular Biology

Year
2
Academic year
2024-2025
Code
01004563
Subject Area
Biochemistry
Language of Instruction
Portuguese
Mode of Delivery
Face-to-face
Duration
SEMESTRIAL
ECTS Credits
6.0
Type
Compulsory
Level
1st Cycle Studies

Recommended Prerequisites

 Molecular Biology or attending those classes.

Teaching Methods

Exposure of the theoretical subject of each class followed by the laboratorial project. On this work students are divided into small working groups where they perform  the practical work, based on Molecular Biology tenhniques and following a given experimental procedure. The methodology used is discussed with the students during the experimental work. By the end, the results are discussed and the students are assigned to write an experimental  report of the subject. Bioinformatic classes have a theoretical and practical component. After all the laboratory classes, students are committed to develop a small project with all the acquired knowledge/techniques and expose it to the classroom mates. 

Learning Outcomes

The main objective of this unit is the acquisition of technical and practical knowledge of the main technologies used in Molecular Biology, as well as the basic concepts of bioinformatics. Therefore, students should develop know how in several techiques such as PCR, genotyping, DNA clonning, resctriction analysis, recombinant protein expression and purification in an heterologous system. Afterwards, students should be taught in bioinformatics, taking advantage of the web available tools and information.

Work Placement(s)

No

Syllabus

Analysis of DNA sequences and genotyping .

PCR amplification of a DNA fragment and analysis of PCR products by electrophoresis.

Cloning: Ligation of a PCR product to a cloning vector; Competent cells preparation and transformation; Screening of the recombinant clones (blue and white screening); Plasmid DNA isolation and restriction digestion.

Expression of a recombinant protein fused and its purification. Analysis of the process by SDS-PAGE followed by Coomassie blue staining and Western blotting.

Bioinformatics: applications and main goals. Basics on databases such as BLAST, PFAM, InterPro.

Head Lecturer(s)

Anabela Pinto Rolo

Assessment Methods

Assessment
Project: 30.0%
Two independent reports, 15% each but students are required 7.5% in each report for approval: 30.0%
Exam: 40.0%

Bibliography

T.A. Brown (2006) Gene Cloning and DNA Analysis, 5th ed., Wiley Blackwell; ISBN: 1405111216.

F.M. Ausubel, R. Brent, R.E.  Kingston, D.D. Moore, J.G. Seidman, J.A. Smith; K. Struhl eds. (2003)l Current Protocols in Molecular Biology, John Wiley & Sons, Inc.

M.R. Green and J. Sambrook (2012) Molecular Cloning: A laboratory Manual, 4th ed., Cold Spring Harbor Laboratory Press; New York

H. Lodish, A. Berk, C.A. Kaiser, M. Krieger, M.P. Scott, A. Bretscher, H. Ploegh, P.T. Matsudaira (2007) Molecular Cell Biology, 6th ed.,  W.H. Freeman and Company