TOPICS AND PROGRAM OUTLINE
The course
This five-day course will be dedicated to fundamental Molecular Biology techniques in DNA, RNA and protein isolation and manipulation. Through theoretical lectures and practical sessions it aims to cover: different DNA cloning strategies, PCR, RNA isolation, RT-PCR, SDS-PAGE, methods for gene expression and protein analysis, and bioinformatics.
The theory and practical aspects of the course are designed so that no previous experience in Molecular Biology is required. Every practical session will be introduced by a lecture. The practical session will be concluded by a question session, when participants will be able to seek clarifications and more detailed explanations about the daily practical. During the practical sessions the participants will work in small groups, assisted by the external international faculty in collaboration with local scientists.
A key aspect will be to contextualize these molecular biology techniques in terms of their applicability for local science and medicine. Hence, the attendants will be taught about how molecular biology is currently applied to medicine and crop science, and how this discipline contributed to the establishment of the CRISPR-based gene editing strategy and of the multidisciplinary synthetic biology approach.
Participants will be invited to present their own projects and to discuss, during a roundtable session, how they would apply the Molecular Biology techniques of this workshop to their own research work.
Finally, there will be a science policy lecture and a roundtable discussion to promote the culture of science advocacy, emphasizing the societal importance of research and addressing scientific funding opportunities in Africa.
Program outline
Day 1 - 21 November 2016
9:00-10:00 Welcome introduction
10:00-11:00 Lecture: macromolecules and biological processes
11:00-11:40 Lecture: Introduction to PCR and experimental design
11:40-12:30 Lab tour and safety instructions
12:30-13:30 Lunch break
13:30-14:30 Practical: Setting up PCR reactions
14:30-15:30 Lecture: DNA cloning: strategies and vectors
15:30 -18:00 Practical: in silico cloning using open access resources
18:00:18:30 Practical: inoculation of liquid E.coli culture for competent cells production
18:30-19:00 Question session
Day 2 - 22 November 2016
9:00-12:30 Practical: preparation of E.coli competent cells, gel purification of PCR-amplified DNA fragments
12:30-13:30 Lunch break
13:30-15:00 Practical: completing preparation of E.coli competent cells
15:00-18:30 Practical: DNA cloning by traditional and Gateway strategy
18:30-19:00 Question session
Day 3 - 23 November 2016
9:00-10:30 Lecture: analysis of gene expression, genome-wide and gene-specific approaches
10:30-12:30 Lecture: experimental approaches of gene expression modification in plant and animal science; gene editing by the CRISPR-Cas9 technique; synthetic biology application in medicine and crop science
12:30-13:30 Lunch break
13:30-14:30 Practical: Transformation of E.coli competent cells
14:30-18:30 Practical: RNA extraction from plants and mammalian cells, RNA quality assessment by gel electrophoresis, RT-PCR
18:30-19:00 Question session
Day 4 - 24 November 2016
9:00-11:00 Lecture: studying proteins, genome-wide and gene-specific approaches
11:00-12:30 Lecture: application of DNA cloning for the production of recombinant proteins and in vitro studies
12:30-13:30 Lunch break
13:30-14:00 Practical: preparation of liquid cultures of selected bacterial clones
14:00-18:30 Practical: soluble protein purification from plant and animal samples; protein quantification by Bradford method; SDS-PAGE running and gel staining by Coomassie Blue staining
18:30-19:00 Question session
20:00-22:00 Social dinner
Day 5 - 25 November 2016
9:00-9:30 Practical: SDS-PAGE gel destaining
9:30-12:30 Practical: plasmid purification from bacterial cultures; plasmid analysis by PCR and restriction digestion
12:30-13:30 Lunch break
13:30-16:00 Roundtable discussion on attendants’ projects and Molecular Biology as an additional approach of study
16:00 -17:30 Lecture: science advocacy and science policy
17:30-18:00 Concluding remarks