Safety
Safety was the most important thing when we engaged in our project. we must follow the rules strictly.
When we began to do our experiments, we have already received a two-week long training about lab safety and common microbiology techniques. After the training, we have learned about basic safety rules and common microbiology techniques in the lab, we could use centrifuge properly and clean bench proficiently. Moreover, when we have any problem about the use of lab equipment, we should ask our instructors for help to avoid being hurt in the lab. We must respect and abide by the laboratory safety rules for the security and health of others and the environment.
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1. Be sure to read all safety instructions and know what to do in case of emergency or accidents.
1. Must wear safety goggles, if any chemicals, heat, or glassware are used.
1. Use the alcohol burner safely, especially when using 75% alcohol on the side.
1. Any container must be labeled indicating its contents and effective time.
1. Carefully read the operating procedures and be trained before using the equipment.
Our project is divided into two parts: molecular biology experiment and cell biology experiment. There are a number of security issues to be aware of in both parts. We have described above the relevant security measures taken all through summer to minimize the risk of incidents in the lab.
In the molecular biology experiment part, one of the main concerns is the dissemination of our engineered strain, which could be a threat to the public and the environment. In the experiment we used E. coli DH5α competent cells. DH5α is a mutagenized strain that mainly exhibits immunological deficiency of foreign DNA. It is a strain used for genetic engineering and lacks a certain degree of immune mechanism compared to normal strains. The DH5α strain does not have antibiotic resistance unless you transform an antibiotic resistance gene into it. E. coli does not cause disease in normal habitats. Only a small number of specific strains can cause inflammation when they enter the organ, and antibiotics are generally effective in treating them. In spite of this, we should clean and autoclave the experimental equipment with disinfectant after the experiment, and specially treat the discarded bacterial liquid to prevent it from polluting the environment. And all the garbage will be classified, unified by the school treatment. In addition, the nucleic acid dyes used in the agarose gel electrophoresis are toxic which could embed in double-stranded DNA, so the electrophoresis area should be classified as a contaminated area, away from the normal test area. And it needs to be cleaned regularly. The cell line used in our cell biology experiments was human embryonic kidney cell HEK-293T. The reagents used in the experiments are non-toxic and pose no risk of diffusion, so the experiments can be performed in a conventional cell culture room. Because the cells are frozen and stored in the liquid nitrogen tank, the temperature of the liquid nitrogen is -196 degrees Celsius, we should wear protective gloves and protective glasses when taking the cells to prevent frostbite or the explosion of the frozen storage tube. We have to wear protective clothing and take precautions when we go into the culture room. There is a buffer room between the cells, which is negative pressure, can ensure the isolation between the outside air and the operation, prevent pollution leakage, and ensure a relatively sterile environment inside. In addition, the most important equipment is the biosafety cabinet, the bioclean safety cabinet can provide dual protection for samples and workers. The filtered clean air flow is blown down from the top of the safety cabinet and passes through the work area, where it is captured before reaching the breathing area of the staff. The air stream will be filtered before it is discharged. Normally, the filtered air will be discharged back to the laboratory or out through the exhaust pipe. Follow the typical layout of "clean to dirty" work in the biosafety cabinet. The left side is used to store clean items, we can be operated in the central area, and the right side is used to store contaminated appliances and experimental materials. We culture the cells in a cell incubator, which provides a 37.5℃, 5% carbon dioxide environment. Waste cells and cell cultures also need to be treated before being discarded. [1] https://www.mcrsafety.com/blog/lab-safety
2. Do not eat or drink in the laboratory or use equipment as containers for food or beverages.
3. Wear proper lab clothing.
4. Ensure to know the locations of all the equipment and materials.
5. The drugs, samples, and solutions used should be labeled. Never load items in containers that do not match the label.
6. Keep aisles clear.
7. Dispose of all the waste properly.
8. Wash hands using soap and water after the experiment.
Clothing
2. Dress in the lab coat and wear gloves.
3. Long hair must be tied back.
4. Dangling jewelry and loose clothing are not permitted.
5. Shoes must completely cover the foot.
Heat
2. Flames should not be used in laboratories using flammable gases and flammable reagents.
3. No smoking.
Chemical substances
2. When using low boiling point organic solvents, be sure to stay away from fire and heat sources. Reagent bottles should be tightly sealed and stored in a cool place.
3. Be familiar with the dangers of the chemical before using it.
Equipment using
2. The equipment and aisles should be kept clean during the process of sample examination.
3. Immediately report the equipment failure to the instructor and should not be handled without authorization.
4. Use water and electricity according to operating procedures.
5. When leaving the laboratory, check whether the instruments, water, electricity, doors, and windows are closed.
Safety for our project
