There is risk associated with almost any laboratory experiment, especially when working with live biological agents. However, our team has gone to great lengths to minimize the risk posed to the researcher, the public, and the environment. Overall, our project can be considered low risk since the finished product is not intended to come in contact with humans in any form. Our E. coli and Rhodobacter sphaeroides cultures should only be grown and tested in a bioreactor with proper laboratory safety technique. Even in the event that either strain was ingested, its is very likely that no harm would occur since the bacteria are not able to survive the environment of the human digestive system. The two major safety concerns associated with our project arise during DNA purification using gel electrophoresis and extraction of PCB from spirulina powder.
Gel electrophoresis is necessary to purify DNA for almost all steps of our project. To make DNA visible as it moves through the agarose gel, Ethidium Bromide (EtBr) is added to the gel to act as a fluorescent tag. Ethidium Bromide itself is a potent mutagen and a known carcinogen that may be absorbed through the skin. For this reason, a separate lab bench has been set aside for all items coming into contact with the toxin including pipets, tips, gel rigs, glassware, and DC power sources (for electrophoresis). In addition there is a vessel designated as contaminated where all disposables (i.e. pipet tips) maybe deposited and properly disposed of at a later date. It is important that all items coming in contact with EtBr remain on the EtBr lab bench and all noncontaminated lab materials are not brought to this bench. Nitrile gloves must be worn when handling EtBr and replaced frequently, especially when going back and forth from this bench to another one.
Phycocyanobilin (PCB) is necessary for the final testing and characterization of our strain of Rhodobacter sphaeroides and is most easily obtained by purifying it from spirulina, a cyanobacteria.
2008 Safety Outline:
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