Team:ZJUT-China/Safety
This year, we successfully continued the good record set by our predecessors and put the safety of ourselves and others at the forefront of our program goals. Ever since the start of our project, safety has always been a priority to the ZJUT-China iGEM team. Before we started, we were trained in safety rules by our teachers in great detail. Only when we passed the exams about lab safety，were we allowed to conduct our experiments in the lab.[1] So，our experiments were designed and operated strictly according to the laboratory safety standards, which includes strict laboratory safety criteria and，most importantly，the safety of bacteria usage during our experiments. The rigorous experimental plans and scrutinous work ethics in the lab cause no harm to the living creatures and the environment in which they live. Dr. Zhang Yinjun is the supervisor of our lab, who is responsible for our lab's security. Before we start our experiments, all our experimental plans were presented to Dr. Zhang Yinjun and Dean’s office in detail. Furthermore, the bio-lab safety officer supervised us frequently, while giving us safety training courses in the laboratory from time to time.
In all laboratories, some basic safety principles must be applied:
- Everyone should mark his/her own things (name + number + date + name) on time and be responsible for his/her own things.
- It is forbidden to drink and eat in the lab.
- It is compulsory to wear appropriate individual personal equipment.
- Long hair must be tied back.
- We must wash our hands regularly.
- The basic clothing: A conventional lab coat, pants and closed shoes.
- When dealing with toxic reagent, be careful not to touch them directly especially when there is open wound.
- If the glassware is broken accidentally, you must be careful with the disposal of fragments in case of cutting.
- The pipette gun should be adjusted back to the maximum range after using.
- When your hands are wet do not touch any electric devices or plugs.
- After the experiment, the glassware should be cleaned and placed neatly, and the used stuff should be returned to their original places.
- Classify the laboratory waste before deposit it.
- The last person to leave the laboratory must check the equipment, lights and doors.
Our laboratory has equipment which meets primary and secondary biosafety standards[2]. And the instruments and equipments used in this project include Centrifuges, Electrophoresis System, Spectrophotometer, PCR Amplifier, ELIASA, Chromatographic columns, Shaker, Vertical Clean Benches, Electric Sterilizer, AKTA Explorer, Constant temperature incubator, Freeze Dryer. All the instruments and equipment should be checked regularly, maintained in time and cleaned before and after the experiment to ensure the accuracy and reliability of the experimental results. Wet lab members who participates in the experiment will receive appropriate training before using the instruments and they will also learn the correct operation of the instruments according to the instructions. Any experiment will be carried out with at least one person present. For the instruments commonly used in the laboratory, the team members sorted out and listed the notes, some of which are shown as follows.
Vertical Clean Benches:
- Don't do experiments when the ultraviolet lamp is on.
- Keep the ignition source away from the ethanol to prevent an explosion while using the alcohol lamp and alcohol.
- Don't push the windshield up and down so hard in case of hurting someone.
Shaker:
- Check whether the base is firm to prevent the lask from toppling, which may cause rust, leakage, and fire.
- Set the temperature of the shaker so that it's not too high to cause fire or too low to affect the propagation of bacteria.
- If there is any abnormal condition (burning smell, etc. ), the power plug should be removed immediately to turn off the instrument, or the power supply should be disconnected by the circuit breaker.
Electric Sterilizer:
- Check whether the base of the shaker is firm, and prevent the shaker from tipping over during shaking, which may cause rust, leakage, and fire.
- Don't touch the upper cover of the sterilizer to prevent skin burn when using.
- When using the electric sterilizer, please be sure to wait until the pressure drops to 0 before opening the exhaust valve, so as not to cause the explosion.
- During sterilization, pay attention to the temperature, pressure and time.
- After sterilization, be careful of the overflow of the steam when opening the sterilization pot to avoid being scalded by steam.
Freeze Dryer:
- Remember to precool for 30 minutes or more before using it.
- Samples prepared by freeze dryer should enlarge surface as much as possible, and should not contain acid, alkali and volatile organic solvents.
As we all know, laboratory safety is not only related to our life safety and environmental safety, but also plays a decisive role in the success or failure of our project. Experimental operation safety is an important part of experimental safety.
- Do not turn on the UV light when operating in vertical clean bench.
- Disinfect and wipe the working area with 70% ethanol or 10% fresh bleach, especially the area that may contaminated with bacteria, before and after the experiment.
- When using an alcohol lamp, do not ignite alcohol lamp with another alcohol lamp; When adding alcohol, be careful not to be less than 1/3 of the volume, not more than 2/3 of the volume; Extinguish alcohol lamp timely before stepping away from the alcoholic lamp.
- When using pipette to draw liquid, pay attention to replacing the nozzle to avoid liquid cross contamination.
- The coating stick or inoculation ring should be burned on the alcohol lamp as sterilization before and after coating or inoculation.
- Wear gloves when using the dye EB during agarose gel electrophoresis and do not contact it directly. The electrophoresis tank should be covered to prevent evaporation of liquid and reduce the possibility of electric shock.
- Wear gloves when using microwave to heat things in case of scalding.
- When using the centrifuge, pay attention to balancing the centrifuge tube by weighing balance, cover the centrifuge tuber, and do not open the hatch until the centrifuge stops completely.
- Before using the autoclave sterilization pot, check whether the amount of distilled water in the pot is sufficient and check whether each valve and switch are normal; When sterilization is finished, cool it naturally before opening the exhaust valve for exhaust.
This year we focused on detecting RNA in the Cell-Free system. We designed our project based on the official requirements of iGEM rigorously and followed the rules strictly.
For the safety of the experiments, non-pathogenic strains were selected, including E. coli KL740 and E. coli DH5α.[3] These chassis microorganisms are commonly used in the laboratory and cleaned after high-temperature sterilization without any features that may endanger human and environmental safety. So, from the point of view of the strains we use, our project is safe.
The plasmids we use in our project: P70a-σ28, P28-tetR, P28-tetR-ssrA, P28-tetO-deGFP, etc. are almost harmless to humans.
In carrying out our human practices work and considering the implementation of our projects, we also consider many issues regarding the unfolding of our projects, i.e. our ethical responsibilities to our audiences, the users of our project products, the environment during our participation activities.
- Wear a mask at all times on buses or subways to protect members from themselves.
- Wear a mask throughout the journey in a taxi.
- If riding out, wear a mask and helmet throughout the ride.
- Obey traffic rules and be careful when driving, cycling or walking.
- Cover coughs and sneezes with tissues or sleeves.
- Reduce exposure to the public in public places.
- Avoid touching face with hands when you are out.
- In crowded places, keep a safe distance of more than one metre between people.
- Try not to eat outside. Avoid direct contact with cutlery when eating with other people.
- Follow the campus vaccination rules and daily self-temperature measurements.
- Wash your hands to a strict standard when you return home from outside.
- If you have been to or come into contact with people in high-risk areas, report it promptly and take care of your own safety and take protective measures.
Besides biosafety and biosecurity, ethical considerations also need to be considered when developing a detection device. Our experiments are designed to be ethical and participation is voluntary for all participants.