Cell and Molecular Biology Experiences overview
Name | Summary | Concepts Covered | Time |
DNA restriction & electrophoresis | Participants perform a restriction digest on a plasmid containing an inserted gene. They run the resulting digest on an agarose gel and use the results to determine whether the gene had been insert correctly or incorrectly. | Structure of DNA, restriction endonucleases and restriction digests, enzymes, cloning using commercial plasmid vectors, demonstration of DNA using agarose gel electrophoresis |
1 day (4.5hrs) or by arrangement |
The polymerase chain reaction | Participants use the polymerase chain reaction to amplify a section of a gene. The presence of this gene fragment is then demonstrated using agarose gel electrophoresis. | Structure of DNA, DNA replication, PCR, enzymes, demonstration of DNA using agarose gel electrophroesis |
1 day (4.5hrs) or by arrangement |
Alakaline lysis mini-plasmid preparations | Participants are provided with a culture of bacteria which has been transformed using a commercial plasmid vector containing a gene of interest. They use an alkaline lysis mini-plasmid preparation to recover the plasmid from the bacteria, followed by a restriction digest and agarose gel electrophoresis to demonstrate the presence of the plasmid and inserted gene in their extract. | Structure of DNA, using plasmids for cloning, genetic modification, microbiology, alkaline lysis mini-preps, restriction endonucleases and restriction digests, enzymes, demonstration of DNA using agarose gel electrophoresis |
1 day (4.5hrs) or by arrangement |
Mitosis movies | Participants prepare a culture flask of mammalian cancer cells. These cells are treated with a bank of chemotherapeutic drugs which are known to affect the cells by interfering with the cell cycle. The flasks are then set up on a microscope which takes a series of images over sixteen hours. By merging these images into a “movie”, participants observe the progression of their cells throughout the cell cycle and try to determine which drug was placed in each well, depending on their knowledge of how each drug is known to work. | The structure of the cell, the cell cycle, mitosis, cell division, cancer and how it is linked to disturbances in the cell cycle, action of chemotherapeutic drugs |
1 day (4.5hrs) or by arrangement |
Immunofluorescence | Participants are provided with coverslips containing cultured mammalian cells. They carry out an immunofluorescence protocol aimed at demonstrating specific cellular structures (eg. actin or tubulin) with a DAPI ounterstain and observe and photograph these cells using fluorescent and / or confocal microscopy | Cell structure, mammalian cell culture, mitosis and the cell cycle, antigen-antibody interactions, direct and indirect immunofluorescence, fluorescence and confocal microscopy |
1 day (4.5hrs) or by arrangement |
Green Fluorescent Protein (GFP) purification and electrophoresis | Participants extract GFP from genetically modified bacteria. They use chromatography to purify the GFP protein and analyse it by SDS-polyacrylamide electrophoresis. | How amino acid sequence determines protein structure, importance of 3D structure of proteins to their function, effects of temperature on protein structure, size determination of proteins |
1 or 2 days |