The Molecular and Genomic Workshop is designed to teach the techniques associated with modern cloning (Recombinant DNA Technology), transformation and analysis of genetic material. The program consists of approximately twenty (20) laboratory sessions.
The Molecular and Genomic Workshop consists of the following components:
1. Biotechnology Laboratory and Safe Practices
Consistent with our aims to provide biotechnology training of international standard; the training programs begin with training in the standard requirements of working in modern biotechnology laboratories. Students are give training in safe working practices in a biotechnology working environment. This session is meant to help students understand how to conduct themselves in modern biotechnology laboratories and lay the foundation for subsequent lab sessions.
2. Recombinant DNA Technology
Recombinant DNA is DNA that has been created artificially, i.e. by human manipulation. DNA from two or more sources is incorporated into a single recombinant molecule. Cloning is the term used for producing many identical copies of the same recombinant molecule. In this section, students will carry out the “construction of a recombinant gene” followed by “cloning”. The following techniques and experiments will be used to achieve those objectives.
I. Bacterial Growth Studies
Teaches the importance of aseptic techniques used for the growth of bacteria. Using aseptic techniques, students will grow bacteria for use in cloning. These bacteria provide the starting material for cloning in the form of plasmids containing agene of interest and a vector.
II. Plasmid Isolation (Alkaline Lysis)
Teaches how to extract plasmids, the small pieces of circular transferable DNA material, from the bacteria. A lab activity for the isolation of extra chromosomal DNA commonly knows as plasmid DNA. Uses a simple alkaline lysis method and alcohol precipitation.
III. DNA Restriction Digestion Analysis
A lab activity that teaches how various restriction enzymes cut DNA sequences at specific location. This session teaches the use of DNA restriction in construction and analysis of recombinant DNA and testing similarity and dissimilarity in two or more genetic materials such as identity and paternity testing.
IV. Purification of a Gene
A lab activity that teaches purification of a gene using agarose electrophoresis followed by gene or DNA elution and purification from the agarose gel: This purification technique can also be used for cleaning DNA or gene preparations of unwanted salts, dyes, and so forth.
V. DNA Ligation
Following restriction enzyme digestion of DNA molecules, researchers need to rejoin the “sticky” and/or “blunt” ends of the DNA to generate recombinant DNA, a process known as ligation. The DNA Ligation lab activity teaches students about ligation through both sticky ends and blunt ends as they ligate DNA fragments together to make a viable plasmid.
VI. Bacterial Transformation
Teaches transformation of bacteria by infusion of genetic materials into the bacteria. A technique widely used for production of recombinant products, includes preparing competent cells, infusion of plasmid followed by selection of transformed bacteria.
VII. Screening for Recombinants
In this lab activity, the importance of using correct screening methodology after the cloning experiment will be emphasized by allowing trainees to screen their transformants.
3. Bacterial Recombination & Gene Expression Experiments
I. Bacterial Conjugation
A lab activity that teaches transfer of genetic material between two bacteria. A process by which genetic materials are exchanged in nature and results in transformation of bacteria.
II. Viral Transduction
This lab activity teaches how to use viruses for introducing genomic information into bacteria. Teaches students about the life cycle of bacterial viruses (phages) and how to manipulate it to introduce genomic information into the bacteria. Students will learn what a plaque is and how to differentiate between them and bacterial colonies, and learn about the lytic and lysogenic phases of transduction.
III. Expression of a Recombinant Protein
Following molecular cloning of a gene into a plasmid, the final stage is the expression of the cloned gene to produce large amounts of protein. Students learn about essential promoters and other elements necessary for successful protein expression in bacteria, including the differences between inducible and constitutive expression. Molecular weight of the expressed protein will be determined by SDS-PAGE.
IV. Correction of Genetic Defect
A major goal of genetic engineering is to have the ability to correct genetic defects to treat genetic diseases, such as cystic fibrosis, sickle cell anemia and Huntington’s disease. This lab activity teaches how to correct a genetic defect in bacteria. Students study for a defective gene in bacteria and their correction by introducing the correct gene. This technique is widely known as bacterial Complementation.
4. PCR & Applications
A lab activity that teaches how a single copy of DNA or gene can be rapidly multiplied into hundreds and thousands of identical gene copies within a short period of time. The process is widely known as Polymerase Chain Reaction or “PCR”. The PCR method is widely used for amplification of specific DNA sequence either for gene cloning, forensic, identity, or paternity testing.
I. Polymerase Chain Reaction (PCR)
Teaches students about the importance of different reagents and the ways to handle them for good amplification. Genomic DNA will be isolated and a specific gene will be amplified by students using PCR and run on a gel to be visualized.
II. Mutation Detection and Analysis
Teaches the concept of point mutation in genetic materials and it phenotypic consequences. Students learn various methods of detection and analysis of mutation and perform a PCR and restrict analysis based detection of mutation. Students learn various human diseases where point mutations play a role and the significance of mutation analysis in clinical diagnostics.