Team:ECNUAS/Safety
Safety
Team lab safety
· Wear protective clothing and disposable gloves
· Tie loose hair up
· Understand proper procedures for handling experimental materials
· Before the experiment, check whether the instrument is intact and the device is correct
· Disinfect the surfaces of all appliances with 70% ethanol before work
· The centrifuge shall be balanced and confirm whether the cover is tight
· Pipette is only used one gear when pipetting, and the second gear is used when the remaining liquid is produced
· Turn off the ULTRAVIOLET rays and turn on the exhaust fan and light when using the ultra-clean platform
· It is not allowed to mix various chemical reagents randomly to avoid accidents
· No eating, drinking, or smoking in the laboratory
· Poisonous drugs shall not enter the mouth or contact the wound
· All reagents should not be taken out of the laboratory
· The remaining waste liquid shall not be dumped into the sewer at will. It shall be poured into the waste liquid tank for recycling
The project safety
Overview
Given the current worldwide challenges in the diagnosis of diabetes, SNA iGEM is establishing a dual-fluorescent reporter screening platform targeting GR lead compounds against type 2 diabetes to reduce the time and effort required to identify the efficacy of GR drugs.
GR is a transcription factor that ACTS on the binding sites of transcription factors to regulate the expression of gluconeogenesis and thereby improve blood glucose concentration, with little risk.
DH5 E. coli is a bacterium that cannot be put into the living environment and can only be grown in a laboratory. The potential health and environmental hazards associated with DH5 E. coli are very limited, so the risk is close to zero.
The traditional EB (EtBr) nucleic acid dye is volatile, toxic, penetrates cell membranes, and induces gene mutations. The YeaRed nucleic acid dye we used is a unique oily large molecule that cannot penetrate the cell membrane into the body and is non-volatile and is a safe non-toxic nucleic acid dye.
Use
of pipette
1.
Pipette tip should be used, and it is strictly prohibited to use the pipette to draw directly.
2. All pipettes shall be fitted with cotton plugs to reduce pipette contamination.
3. Do not blow gas into solutions containing infectious substances.
4. Infectious substances cannot be repeatedly blown and mixed with the pipette.
5. The liquid cannot be blown out forcefully from the pipette. 6. The mark-to-mark pipette does not need to discharge the last drop of liquid, so it is better to use this pipette.
7. Contaminated pipette should be completely immersed in a crush-proof container with appropriate disinfectant. The pipette should be soaked in disinfectant for an appropriate time before handling it.
8. Containers for the discarded pipette should not be placed outside but should be placed in the biosafety cabinet.
9. Syringes with fixed hypodermic needles cannot be used for pipetting.
10. When opening diaphragms in bottles, use tools that can use pipettes instead of hypodermic needles and syringes.
11. To prevent the spread of infectious substances from dripping from the pipette, a piece of cloth soaked in disinfectant or paper soaked in disinfectant should be placed on the worktable and treated as infectious waste after use. 12. The contaminated pipette shall be soaked in an appropriate disinfectant and sterilized in a high-pressure disinfection pot, then washed with tap water and rinsed with deionized water.
Use
of biosafety cabinets
1. National
standards and relevant literature should be referred to to introduce the use methods and limitations of biosafety
cabinets to all potential users. Written rules, safety manuals or operation manuals shall be issued to the staff.
In particular, the safety cabinet is no longer able to protect the operator in the event of overflow, breakage, or
poor operation.
2. Biosafety cabinet can only be used when it is running normally.
3. The glass baffle shall not be opened in the biosafety cabinet during use.
4. Equipment or specimens should be placed as little as possible in the safety cabinet, so as not to affect the airflow circulation of the rear pressure exhaust system.
5. Bunsen lamps should not be used in the safety cabinet, otherwise the heat generated by combustion will interfere with the airflow and may damage the filter. Micro electric heaters are allowed, but disposable sterile inoculation rings are preferred.
6.
All work must be done in the middle and back of the workbench and can be seen through the glass observation
baffle.
7. Minimize the movement of
personnel behind the operator. 8. The operator should not repeatedly move out and extend the arm to avoid
interference with the airflow.
9. Do not block the air grid with laboratory notebooks, pipettes and other items, as this will interfere with the flow of gas and cause potential contamination of the items and operator exposure.
10. Wipe the surface of the biosafety cabinet with appropriate disinfectant after work and before leaving work every day.
11. Before and after the work in the safety cabinet begins, the fan in the safety cabinet should run for at least 5 minutes.
12. When operating in the biosafety cabinet, it is not allowed to carry out writing work.
13. Through the artificial generation of microorganisms (bacillus subtilis) in the biosafety cabinet, the collection of bacillus subtilis spores in the culture plate outside the biosafety cabinet can be used to determine the protection ability of the biosafety cabinet for the staff.
Use
of centrifuges
1. In the use of laboratory
centrifuges, good mechanical properties of instruments are the prerequisite to ensure the safety of
microorganisms.
2. The centrifuge shall be operated in accordance with the operation manual.
3. The height of the centrifuge should enable the small staff to see the inside of the centrifuge, so as to place the cross shaft and the centrifugal barrel correctly.
4. Centrifuge tubes and containers for centrifuge specimens shall be made of thick-walled glass, or preferably plastic, and shall be inspected for damage prior to use.
5. Test tubes and specimen containers used for centrifugation should always be tightly closed (screw caps are preferred).
6. Loading, balancing, sealing and opening of the centrifugal barrel must be carried out in the biosafety cabinet.
7. The centrifugal barrel and the cross shaft shall be paired according to weight and balanced correctly after loading the centrifugal tube.
8. The operation guide shall provide the space to be set aside between the liquid level and the orifice of the core tube.
9. Empty centrifugal barrels should be balanced with distilled water or ethanol (isopropyl alcohol, 70%). Salt or hypochlorite solutions are corrosive to metals and therefore cannot be used.
10. For grade 3 and 4 microorganisms, a sealable centrifuge bucket (safety cup) must be used.
11. When using a fixed-angle centrifugal rotor, care must be taken not to overfill the centrifuge tube, otherwise leakage will result.
12. The cavity walls of the inner rotor of the centrifuge should be checked daily for contamination or contamination. If contamination is apparent, the centrifugal operation specification should be reassessed.
13. The centrifugal rotor and the centrifugal barrel should be inspected daily for corrosion or minor cracks.
14. After each use, clean up the pollution of the centrifuge barrel, rotor and centrifuge chamber. 15. After use, the centrifugal barrel should be placed upside down to dry the balance liquid.
16. When a centrifuge is used, infectious particles which can be transmitted through the air may be ejected. If the centrifuge placed in front of the traditional open Ⅰ level or Ⅱ level in biological safety cabinet, the particles cannot be due to too fast safe incase of air entrapment. And in centrifugal closed Ⅲ level in biological safety cabinet, can prevent the generated aerosols are widely spread. However, good centrifuge techniques and tightly capped centrifuge tubes provide adequate protection against the production of infectious aerosols and diffusible particles.
Use
of homogenizer, shaker, agitator and ultrasonic processor
1.
Household (kitchen) homogenizers cannot be used in the laboratory as they may leak or release aerosols. It is
safer to use special laboratory agitators and digesters.
2. The lid, cup or bottle should be kept in normal condition without cracks or deformation. Cover should be able to seal tightly, liner should also be in a normal state.
3. When homogenizer, shaker and ultrasonic processor are used, pressure will be generated inside the container, and aerosols containing infectious substances may escape from the gap between the lid and the container. Plastic containers, especially Polytetrafluoroethylene (PTFE), are recommended because glass can break and release infectious material that can harm the operator.
4. When using the homogenizer, shaker and ultrasonic processor, the equipment should be covered with a strong and transparent plastic box and disinfected after use. If possible, these instruments can be operated in a biosafety cabinet covered with a plastic cover.
5. Open the container in the biosafety cabinet after the operation.
6.
Hearing protection shall be provided to personnel using ultrasonic processors.
Maintain
and use refrigerator, freezer and incubator
1.
Refrigerators, low-temperature refrigerators and dry freezers should be defrosted and cleaned regularly, and all
ampoules broken during storage should be cleared out And test tubes. Thick rubber gloves should be worn during
cleaning and facial protection should be carried out. The inner surface should be disinfected after cleaning.
2. All containers stored in refrigerators should clearly indicate the scientific name of the contents, the date of storage and the family name of the person storing them Name. The unmarked or used articles shall be autoclaved and discarded. 3. A list of frozen items should be kept.
4. Do not place flammable solutions in the refrigerator unless there are explosion-proof measures. This should be marked on the refrigerator door.
5.
The maximum temperature of incubators is 60℃.
Open
the specimen tube and sample
1. The
specimen tube should be opened in the biosafety cabinet.
2. Gloves must be worn, and protection of the eyes and mucous membranes (goggles or masks) is recommended.
3. Wear a plastic apron over protective clothing.
4.
When opening the specimen tube, hold the plug with paper or gauze to prevent spillage.
Automatic
instrument
1. In order to avoid the spread
of droplets and aerosols, these instruments should be of the closed type.
2. The effluent shall be collected in a closed container for further autoclaving or abandonment.
3. Sterilize the instrument according to the operating instructions after each step.
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