Team:Virginia/Safety
Introduction
The safety and security of a project is one of iGEM’s core focuses when developing a device. So, it was only natural that safety became an utmost concern throughout the entire design process and running trials with our device. It was also critical to evaluate every team members knowledge on general lab safety as our team is diverse with members with many different skill sets and abilities. As a result, some team members were experienced in certain lab techniques and able to educate others on safe lab protocol. Through intense scrutiny about our project’s safety and educating each other on proper lab protocol, we were able to devise appropriate security measures.
Training and Enforcement
Prior to beginning any experimentation or working in the lab, the entirety of the Virginia iGEM team completed Biosafety Training courses offered by UVA’s Environmental Health & Safety Department. These instructed team members on critical norms of lab safety such as aseptic technique, autoclave safety, chemical storage and disposal, and hazard assessment. Our lab is Biosafety Level 2 (BSL-2) certified and, as such, we have access to an autoclave, chemical showers, eyewash stations and biological safety cabinets.
Extraction Methods
Our extraction protocol for PH3B involves the use of chloroform, methanol and Sodium Dodecyl Sulfate. Chloroform is an acute category 3 oral, dermal and inhalation toxin. Chloroform is also a category 2 skin and eye irritant. Methanol is a category 3 flammable substance and skin irritant (data from MSDS sheets). When working with these chemicals, all work is done under a fume-hood with gloves to prevent any contact with the skin and any vapor inhalation. Methanol is kept properly stored in a flammable container cabinet and chloroform is kept sealed underneath the fume-hood. These safety precautions will prevent lab members to being exposed to harmful chemicals while extracting P3HB.
Sodium Dodecyl Sulfate is an acute category 4 oral toxin and acute category 3 dermal toxin. SDS is also a category 2 skin and eye irritant. To prevent injury when working with Sodium Dodecyl Sulfate, latex gloves, chemical splash goggles and face masks are worn at all times.
For sonication all lab members in the vicinity must be wearing proper ear protection, which is sound-proof, over the head, earphones that prevent any damage to the ear from noise. This ensures that during the process of sonication, no one’s hearing will be harmed or impaired.
Ensuring Safe Project Design
The chassis for our device is E.coli K-12 TG1. This organism is nonpathogenic (Risk Group 1) and is used in biology laboratories across the country. Aseptic technique is always utilized when handling our chassis organism. Some parts for our genetic device originate from Pseudomonas Putida (sty operon) and Cuprivadus Necator (pha operon). P. putida is a Risk Group 2 organism while C. necator is a Risk Group 1 organism. No check-ins were required for either parts. Both plasmids containing our genetic device have control methods to ensure that the bacterium will not maintain the plasmids under specific conditions. The split kanamycin resistance ensures that the future generations will not survive under antibiotic selection if even one of the plasmids is lost. In addition, limiting the possible escape of the chassis is important as well. The plasmids are costly to maintain without sufficient medium, thus any escape of bacteria will likely cause the loss of the plasmids within a few generations.
