Team:USTC/Safety
Project safety
Our project was designed to be a higher efficient synthesis platform with engineered bacterial applied in factories. There was nothing harmful to the public or environment. However, H2S and CdS, potential products in our photosynthesis system, might have been a risk for us, the researchers and we solved the problem successfully with the help of our universities and the SRB Safety Handbook of the team UNITN-Trento in 2012.
Bacterial strains
We worked with non-pathogenic E.coli strains---Top 10 to amplify our plasmids and BL21(DE3) to express our target proteins in most experiments. To find out whether CdS nanoparticles can generate electrons or not, we used a bacteria called Shewanella oneidensis MR-1, which are non-pathogenic and well-characterized.
Parts
None of the new parts will raise safety issues for further applications except for the parts with the CysDes (BBa_K2242233, BBa_K2242633). We realized these parts would produce hydrogen sulfide at a very low concentration after induced from the wiki of team UNITN-Trento in 2012 at the beginning of our research. We stressed H2S issue again on the main page of the Parts that we submitted. When we did related research about Cysteine Desulfhydrase in our photosynthesis system.
H2S
We knew that H2S, a toxic substance, would be produced at a very low concentrations when Cysteine Desulfhydrase, an important enzyme in our photosynthesis system, was induced. Before experiments started, we read the wiki of the team UNITN-Trento in 2012 and other references to find a solution. We did all the experiments involved in the bacteria producing H2S in specific hoods. We used a separate waste container to dispose of bacteria producing H2S. Eventually, these media and cultures were properly disposed in terms of the way mentioned in our “Protection Guide of lab work”.
As for further application in factories, the problem of tiny amounts of H2S releasing is easy to solve with a well ventilation system and respiratory, skin and eye protection provided.
CdS
Nano CdS can be toxic if not handled correctly. In our experiments, we recovered nano CdS by means of centrifugation and encapsulated it to ensure no direct interaction with environment. Ultimately, the hazardous waste was collected by our schools and disposed properly.
When our project is performed in factories in future, nano CdS can be recovered properly and then applied in other area such as a semiconductor.
Lab safety
We did nothing that would raise any safety concerns for the public or environment. And we took several measures to make sure us, the researchers, work in a completely safe environment.
Training
We did experiments in a well-equipped teaching laboratory (Biosafety Level 1). Before we started experiments in the lab, we received some training from Ms. ZhangQian, one of administrators of our school. And we learned how to use the facility in the lab and how to dispose the chemical, biological wastes and other hazardous wastes such as broken glass. Then we wrote a brief protection guide to warning ourselves about the safety when working in the lab.
In May, our lab delivered a lecture to emphasize the importance of the safety and discussed that how to deal with the potential risk about H2S and CdS when we would express the Cysteine Desulfhydrase.
Figure 1: Scene of the lecture about safety
Protection Guide of lab work
Figure 2: Our protection guide (details are shown as follows)
Click here to download the full version of this handbook!
Personal Protection
1. Lab coats must be worn and fastened until all experiments have been completed and they must be taken off outside the laboratory.
2.Gloves must be worn when doing experiments and they must be removed and properly disposed after experiments. It is strictly prohibited to touch mouse and keyboard of computers, door handle, button in elevator with gloves.
3.When there is a potential risk of splashes or flying particles, goggles must be worn to protect eyes.
4.Suitable shoes and trousers must be worn to protect leg, toes and heels from risky chemical reagents. It is strictly prohibited to wear sandals and shorts when doing experiments containing hazardous chemicals.
5.Tie back long hair when doing experiments.
6.Hands must be washed as soon entering into lab, completing experiments. And everyone must wash hands before leaving lab.
Figure 3: Emphasis the importance of safety
Laboratory Working Area
1.Never eat or drink in working area.
2.Forbid doing other experiments in Specified area. For example, it is strictly prohibited to extract plasmid in the region of running gel.
3.Clean up all spills after work.
4.Be careful when centrifugation: Balance the tubes in the rotor. Do not open the lid while the rotor is moving.
5.Be careful when using autoclave.
6.Be careful when using Bunsen burners. Do not leave Bunsen burners unsupervised and do not use them after hours.
7.When working with UV light during gel extraction, ensure that no skin is exposed to UV.
Reagent Use
1.Never remove any chemical from the laboratory.
2.Do not alter the laboratory set-up.
3.Know the chemical involved an experiment in details. (1) the name of the chemical; (2) hazard presented by the chemical; (3) amount of the chemical use.
Waste disposal
1.Dispose Reagent according to the 《Classification of Hazardous Chemical Wastes by USTC (Trial)》
2.Broken glass should be packaged with paper and plastic bags, labelled "BROKEN GLASS" before being discarded in containers marked "BROKEN GLASS”.
3.Disposal of bacterial cultures: Add 1 to 2 milliliters of thimerosal 84 to each test tube or submerge the plates containing bacteria in the water-thimerosal 84 mixture, then pour the liquid down a drain 15 minutes later.
Figure 4: Containers for different wastes
Reagents
The chemicals we used in experiments that were associated with safety problems include Coomassie brilliant blue G250, ethyl acetate, acetone, ethanol, methanol, glycerol, suflfuric acid, hydrochloric acid, acetic acid, hydrogen peroxide, sodium hydroxide, isopropanol, N, N, N’, N’-tetramethylbenzidine, Ethyl (S)-4-chloro-3-hydroxybutanoate and ethyl 4-chloro-3-oxobutanoate. When we did experiments involved chemicals above, we protected ourselves with lab coats, gloves, goggles worn. After completing experiment, we disposed these waste according to the 《Classification of Hazardous Chemical Wastes by USTC (Trial)》
Figure 5: 《Classification of Hazardous Chemical Wastes by USTC (Trial)》
Shipment safety
We submitted our parts legally and successfully. When sending our parts, we followed the instructions on the Help Submission site: http://parts.igem.org/Help:Submission. First, our samples were sent as miniprepped plasmid DNA (pSB1C3). Second, We submitted parts in 96 well plates, sealed completely (adhesive foil/plastic), and covered with a protective lid. Third, we provided the tracking number of our shipment. Fourth, our samples were not be disguised or hidden to avoid customs. In addition, we consulted with local post offices to ensure less risk as possible.
