Molecular Cell Biology Techniques Level

AY2017/2018 Semester 2

Each practical will have a pre-briefing/ instruction and a de-brief/discussion. Students will be provided with online lab manuals that can be downloaded. In practical #1, students will attend a computer lab to learn about basic bioinformatics concepts, such as generation of database, sequence comparison, substitution matrix. They will also perform blast search and data analyses. In practical #2, the concept of pH and buffers will be reinforced followed by hands-on to prepare solutions and buffers for subsequent practicals. Students will perform SDS-PAGE to resolve and analyze hen egg white proteins. In practical #3, students will perform SDS-PAGE of serum proteins and probe for IgG by Western-blotting followed by enhanced chemilluminescence detection. In practical #4, students will learn aseptic techniques in microbial cell culture and perform bacteria inoculation on agar plate. In practical #5, the concept of using different staining methods to identify microbes will be covered followed by hands-on session. In practical #6, students will perform eukaryote cell counting using haemocytometer and to prepare plant sections for examination under microscope. In practical #7, students will perform immunofluorescence staining of eukaryotic cells and examine the cells under bright-field and fluorescence microscope. In practical #8 & #9, students are required to perform molecular cloning using PCR-based method. The construction of expression plasmid will be used for practical #10 and #11. In practical #10 and #11, students will perform experiments using E.coli to express the recombinant protein followed by purification of this protein.