Methods and Techniques in Molecular Biology

Recommended Prerequisites

Basic knowledge of Biochemistry and Cell Biology.

Teaching Methods

For each of the 5 practical assignments performed throughout the semester a summary protocol will be provided to students through ‘Inforestudante’, which students should read before the class in addition to conducting autonomous research. In the practical classes, students’ autonomy is encouraged for the accomplishment of practical assignments, carried out in groups of 4 students. The majority of practical assignments are carried out in consecutive weeks. Each group of students presents one of the accomplished assignments orally and writes a report on one of the accomplished assignments.

Learning Outcomes

The objective of this curricular unit is thatthe students  learn about the fundamentals and methodologies that are essential for the study of macromolecules with biological importance. Another objective is that students understand the possible applications of the use of those technologies and approach the scientific advances emerging in this field which are the result of technological advances. Students should use their acquired knowledge and competences to improve their knowledge when facing new situations.

 

Competences to be acquired:

- Handle the molecular biology basic laboratory equipment;

- Isolate, analyse and identify biomolecules;

- Carry out biological diagnosis;

- Manipulate genetic material;

- Carry out genetic analysis;

- Carry out bioassays;

- Obtain information, design experiments and interpret the results;

- Elaborate and implement projects in biology;

- Competences to apply the theoretical knowledge in practice;

- Competences in critical thinking.

Work Placement(s)

No

Syllabus

- Preparation of solutions;

- Colorimetric methods to determine quantity and activity of proteins;

- Fluorescence Microscopy: DNA marking, immunocytochemicaland detection of fusion proteins, using Green fluorescent protein;

- Amplification of a DNA fragment by PCR from human genomic DNA;

- Restriction analysis of a DNA fragment amplified to identify a polymorphism (RFLP). Cloning of a DNA fragment amplified by PCR;

- Connection of the PCR-amplified DNA fragment to a cloning vector;

- Bacterial transformation with the recombinant vector;

- Recombinant protein expression in Escherichia coli; purification and analysis of recombinant protein;

- Purification of recombinant protein by affinity chromatography;

- Analysis of recombinant protein by polyacrylamide gel under denaturing conditions.;

- Western blot with a specific antibody against the produced recombinant protein

Head Lecturer(s)

Ana Luísa Monteiro de Carvalho

Assessment Methods

Assessement
Report of a seminar or field trip: 10.0%
Laboratory work or Field work: 25.0%
Exam: 65.0%

Bibliography

WILSON, K. & WALKER, J. (2010) Principles and Techniques of Biochemistry and Molecular Biology. 7ª ed. Cambridge University Press.

BROWN, T. A. (2010) Gene Cloning and DNA analysis. 6 ª ed. Blackwell Science.

VIDEIRA, A. (2001) Engenharia genética: princípios e aplicações. LIDEL.

GLICK, B.R. & PASTERNAK, J.J. (2009) Molecular Biotechnology. 4ª Ed. ASM Press.

WATSON, J.D.; GILMAN, M.; WITKOWSLI, J. [et.al.] (1992) Recombinant DNA. W.H. Freeman and Company. ISBN: 0-7167-1994-0.

BERG, J.M.; TYMOCZKO, J.L.; STRYER, L. (2011) Biochemistry7ª Ed. W.H. Freeman and Company