Safety

Safety of both scientists and the general public is of the utmost importance to anyone working in a laboratory setting. We stringently followed all relevant safety protocols and chose a project that would pose little to no threat to the safety of researchers, the public, and the environment.

Material Safety

Researcher Safety
Working in a laboratory has the potential to put researchers in danger if the proper safety measurements are not followed. To minimize this risk, researchers were trained in safe laboratory practices as described in the Biosafety Provisions section below. The chemicals we used that presented a potential threat to researcher safety were: ethidium bromide, for visualization during gel purification, and SU-8 photoresist and developer, in order to construct the mask used to make our microfluidic device.

• Ethidium bromide: All researchers were required to wear nitrile gloves when handling EtBr. After handling toxic chemicals, researchers were required to discard contaminated items, such as gloves, so as not to spread contamination. Gels containing EtBr were properly discarded in a biohazard bin. When viewing gels, exposure to UV light was minimized through the use of UV-protective safety masks covering the eyes and face. http://www.sciencelab.com/msds.php?msdsId=9927667 MSDS for Ethidium Bromide
• SU-8 developer and photoresist: During construction of our microfluidic mask in the clean room, SU-8 photoresist and SU-8 developer were handled in a ventilated hood. Researchers were required to wear gloves, lab coats, and safety goggles at all times. SU-8 photoresist and developer were discarded in the proper disposal containers. http://cnl.colorado.edu/cnl/images/MSDS/microchem su-8_resist.pdf MSDS for SU-8 resist http://www.ee.iitb.ac.in/~nanoe/msds/su8.pdf MSDS for SU-8 Developer
• Flammable liquids: Flammable chemicals were stored in the flammable closet and handled in a fume hood away from fire. Chemicals were disposed of in the proper receptacles according to Weill Hall's safety requirements. These included: Acetone and IPA, to rinse SU-8 developer, and ethanol, for sterilization and dilution of streptavidin coating reagents.

General safe lab practices were followed in order to ensure researcher safety. Researchers were required to wear nitrile gloves and close-toed shoes during all laboratory work.

Public Safety
It is a common misconception that bacterial strains used in research are dangerous to public health and safety. However, the bacteria we used, DH5a electrocompetent E. coli, are non-pathogenic and unlikely to survive outside of a laboratory setting. To insure minimal risk to public safety, bacteria were killed with 10% bleach before disposal. We perceive no potential threat to public safety from our project. The biochemical pathway we used converts tryptophan, a common amino acid found in many foods, to prodeoxyviolacein, a green pigment. Both chemicals and all intermediates are non-toxic and should present no threat to public health or safety. Furthermore, no gloves were allowed to leave the laboratory, in order to prevent contamination of pubic areas.

Environmental Safety
Our project poses no identifiable threat to environmental safety. As stated above, the DH5a E. coli used in our project is not able to survive outside the lab, and all cells disposed of safely after disinfection with 10% bleach. Bio-hazardous and flammable chemicals were disposed of by following the proper regulations, as stated above. As stated above, no gloves were allowed to leave the laboratory so that chemical and biological hazards were restricted to the laboratory.

Security Risk
We perceive no threat to security from our project. The final product of our biochemical pathway is a harmless pigment which could not be used with malicious intent. While our "biofactory" could be adapted for use in the synthesis of a harmful chemical, the microfluidic device is simply tool to cut costs of synthesis in an industry setting, and does not present any new security threat despite its potential to be misused.

Safety of BioBricks
None of the BioBricks submitted to the registry raised any safety concerns. The parts which we worked with are GFP, RFP, VioA, VioB, and VioE attached to avitags. These BioBricks already exist in the registry, and the addition of the avitag, which allows the molecules to be tethered to our microfluidic device, does not raise safety issues in the use of these parts. Our last BioBrick, the light lysis package, causes cells to lyse in the presence of green light. This should not raise any safety concerns.

Biosafety Provisions

Biosafety Rules and Procedures
We complied with Weill Hall’s safety requirements in gaining access to laboratory space, as well as in use of the laboratory. All safety information and procedures are linked on the main Weill safety page [http://blogs.cornell.edu/whfs/weill-hall-safety-links-and-information/ here].

Weill Hall Safety Personnel
Scott D. Emr is the director of Weill Hall which is the building our lab is located. There is also a Weill Hall Safety Committee. While we did not personally meet as a group with either groups, we met with Dr. Archer, who introduced us to the laboratory space and reviewed safety procedures. Dr. Archer was also present during and helped us with the safe construction of our microfluidic mask.