Team:UPRM/Safety
Laboratory Safety
Safe Project Design
iGEM UPRM’s project attempts to re-engineer the sensing domain of the NarX-NarL two component system in E. coli strain DH5 α by fusing it with the Cph1 photoreceptor domain naturally found in cyanobacteria (Synechocystis sp.). Consequently, E. coli will have the capacity to change from a chemical to photoreceptive activation by responding to a red light and start the production of a byproduct, like macromolecules, by inserting specific genes of production into the multiple cloning site (i/o command). After the gene construct, full expression the reporter gene GFP will be expressed as well proving the functionality of our prototype. To validate it, we will be conducting experiments such as cloning each part, insertion of our genes (transformation) growing our bacteria in cell cultures and conduct different GFP measurement protocols, qualitative and quantitative like absorbance in a Plate reader and flow cytometry. Risks that could arise while conducting these experiments can affect the investigators, the environment, and the experiment itself. The risks are:
- Contamination from E. coli cultures that can lead to unwanted cross contamination from our modified bacteria or external bacteria/ microbes due to poor aseptic techniques.
- Broken glassware that could cut the investigators.
- Antibiotic resistance if antibiotic is exposed to other surfaces like work bench and laminar flow hoods
- When preparing culture mediums, burns from handling hot glassware can happen if protection is not used.
- When working with culture plates and tubes burns can be caused by Bunsen burners if not used properly.
- Unattended Bunsen burners and open gas lines can cause a fire or gas leak in the laboratory.
- UV-Light exposure caused by misuse of the laminar flow hood
- Ethidium bromide exposure when working with agarose gels
- Chloroform and other solvents inhalation or exposure during nucleic acid
Proper precautions will be taken to avoid all types of risks. We believe that minimum risks are involved with our prototype, nevertheless, the application that could be given to it can pose some challenges. For example, since our prototype is a bioproduction “machine”, it could be misused by producing toxic proteins or even antibiotics that if mishandled can lead to resistance in other bacteria. As stated before, we believe that with the scope of our project the risks are limited to laboratory techniques. Nevertheless, as a risk management we will keep our members training up to date. They will attend different conferences that talk about this topic (laboratory safety and techniques).
Safe Lab Work
In our lab, we abide by the general safety guidelines (no eating, drinking, smoking or managing possible contamination sources), laboratory dress code (lab coat, gloves, closed-toe shoes, long pants, tied hair, no exposed skin) and the correct use of laboratory equipment (burners, fume hoods, laminar flow hoods, scales, waterbaths, centrifuges, etc), in which we were properly trained by our PI and graduate advisors.
Unique to our lab:
All members working in the lab are required to complete a mandatory safety training test administered by our PI. This training consists of both a written test, that we are to complete and hand in, followed by a review and discussion of the answers with our advisor as well as guided tours around the safety showers, chemical storage shelves, location of SDS, emergency exits and evacuation routes and disposal areas (biohazard, broken glass containers, chemical waste). No student without training is allowed within the laboratory area and those certified still work under the supervision of a responsible, authorized, graduate student or the PI himself.
Since most of our risks involve laboratory techniques we are constantly being trained in:
- Lab access rules
- Good microbial technique (such as lab practices)
- Disinfection and sterilization
- Chemicals, fire and electrical safety, biosafety levels, personal and protective equipment (including lab coats, gloves, eye protection, etc)
- An inventory control system (measures to track who has what materials and where they are)
- Access controls (measures to control who can access your work spaces, or where materials are kept)
and others. Since we work on a Open bench in a Level 2 laboratory is extremely important for us to keep a sterile and safe environment for our team and our laboratory colleagues.
Thanks to our sponsors
