We used ultraviolet to kill the colonies on the membrane and rupture them to release the laccase. After remaining the bacterial cultivation medium with ruptured bacteria on the membrane for 4 hours, rinse the excess residue with a buffer solution and, then, we receive a membrane embedded with enzyme. This will not cause bacterial leakage. Eventually, the membrane samples taken out of the bacterial laboratory do not carry any bacteria.
For the safety of our project, we employed E.coli DH5α and E.coli BL21 (DE3). These microorganisms are categorized in risk group 1, which means they pose a low risk to human safety and the environment. They have been widely studied and are known for their relatively benign nature, ensuring that our project can be carried out with minimal potential harm.
1. Anyone, including but not limited to staff, managers, relatives and friends, government administrative personnel, personnel from other companies, when working, visiting, inspecting or conducting other activities in the laboratory, must abide by the relevant safety specifications of the laboratory;
2. If internal personnel of the company violate the laboratory safety specifications, they will be punished according to the unsafe behavior level (I, II, III, IV). Unsafe behaviors are classified according to the corresponding regulations of the safety specifications and are marked. Unmarked ones are regarded as (I, II level). Those causing serious consequences are all regarded as level IV. At the same time, they also need to bear all the consequences of the unsafe behavior, including but not limited to property damage, personal accidents, and work delays. When the unsafe behavior violates multiple safety specifications at the same time, it is triggered according to the highest level;
3. Laboratory staff must be very familiar with the relevant usage methods of first aid kits, eyewash devices, emergency showers and laboratory safety specifications. The laboratory supervisor will conduct random safety knowledge checks irregularly every month.
4. Idle people are prohibited from entering the laboratory. Any laboratory staff has the responsibility to refuse irrelevant people from entering the experimental area. Before inspection by government administrative personnel, visits by personnel from other companies or visits by relatives and friends, the laboratory supervisor should be notified and approval is required before entry;
5. Smoking and using open flames are strictly prohibited in the laboratory; (II, III)
6. It is prohibited to charge electronic devices in the experimental area. When using electronic devices, avoid approaching heat sources or any chemical reagents; (II, III)
7. It is prohibited to eat, smoke or store food in containers that may be contaminated in the laboratory; (II, III)
8. Ensure that the labels on the purchased reagents are intact. If the labels fall off, supplement the missing information with label paper; when using bottle A for reagent B, it should be labeled according to the standard label, and the original reagent information should be completely erased and covered on the container to avoid misleading;
9. When preparing reagents for experiments, labels should be affixed. The label content should include the name, concentration, preparer, preparation date and other necessary information;
10. When preparing samples, in addition to the above content, they should be labeled in the format of [abbreviation of name - page number of notebook - number]. After daily use, the samples should be placed in the constant temperature and humidity sample retention room;
11. It is prohibited to take drugs out of the laboratory without permission; (II, III)
12. To ensure that the reagents are not contaminated, the reagents should be taken out of the reagent bottle with a clean ox horn spoon. It is never allowed to grab them by hand; (II, III)
13. Liquid reagents can be poured or sucked with a clean and dry graduated cylinder or straw. The taken reagent cannot be poured back into the original bottle;
14. When opening the stopper of a volatile reagent bottle, do not point the bottle mouth at the face, and do not inhale sharply with the nose close to the reagent bottle mouth. If it is necessary to smell the odor of the reagent, keep the bottle mouth away from the nose and wave your hand above the reagent bottle to allow the air to be drawn towards yourself to smell the odor; (II, III)
15. After taking out the reagent, the stopper should be tightened and the wrong stopper should not be replaced;
16. When disassembling and assembling glass tubes and rubber tubes or rubber stoppers, they should be moistened with water first, and a cotton cloth should be padded on the hand to avoid breaking the glass tube and causing injury; (II, III)
17. When diluting concentrated sulfuric acid, the container should be placed in another plastic container. Only concentrated sulfuric acid can be slowly added to water, not the other way around. If necessary, cool it with water; (II, III)
18. When using alcohol lamps, be careful that the alcohol should not be filled to more than 2/3 of the capacity. When the alcohol in the lamp is less than 1/4 of the capacity, the flame should be extinguished before adding alcohol. The burning lamp flame should be extinguished with a lamp cap and not blown out with the mouth to prevent the alcohol in the lamp from igniting. Alcohol lamps should be ignited with a suitable fire source and should not be ignited with another burning alcohol lamp to prevent fire; (II, III)
19. In case of a local fire, the power supply should be cut off immediately and extinguished by covering it with a wet cloth or asbestos cloth. If the fire is fierce, contact the relevant department immediately and request rescue;
20. Protective equipment should be worn when opening the stoppers of concentrated hydrochloric acid, concentrated nitric acid, and concentrated ammonia water reagent bottles;
21. Corrosion-resistant containers must be selected for storing strong acids, strong bases and corrosive items;
22. When heating flammable solvents, it must be carried out in a water bath to avoid using open flames; Do not put the electrothermal furnace in the laboratory cabinet to avoid fire; When heating or pouring liquids, do not look down at the container to prevent liquid droplets from splashing and causing injury. When heating test tubes, do not point the tube mouth at yourself or others to prevent the medicine from spraying and hurting people; (II, III)
23. When connecting or disassembling glassware with rubber tubes or rubber stoppers, it is best to wrap the glassware with cloth. When connecting, generally hold the instrument to be inserted in the left hand and the inserting instrument in the right hand, and slowly insert it by rotating clockwise. It must be rotated in one direction. Do not point the glass tube mouth at the palm of your hand;
24. After the experiment, conduct a systematic inspection of the laboratory; At the end of work, sort out the equipment, tools and various materials, cut off the power supply, close the doors and windows and faucets.
25. Before the experiment, understand the MSDS of the drugs used;
26. In case of burning by acid or alkali solution, quickly rinse the affected area with a large amount of tap water. If it is a burn by acid solution, rinse with 2% sodium bicarbonate. If it is a burn by alkali solution, rinse with 2% boric acid, and then rinse with clean water. In severe cases, after simple treatment, immediately send to the hospital for examination;
27. Swallowing poison by mistake. The common detoxification method is to give the poisoned person an emetic, such as soap water, water or give them egg white, milk and cooking oil, etc., to ease the irritation. Then use a clean finger to reach into the throat to induce vomiting; (IV)
28. People poisoned by phosphorus cannot drink milk. They can take an oral dose of 5-10 milliliters of 1% copper sulfate solution mixed with a cup of warm boiled water to induce vomiting, and then be sent to the hospital for treatment;
29. Burns or scalds. Do not rinse with water after being burned. You can apply burn ointment on the wound or rub it with concentrated potassium permanganate solution until the skin turns brown, and then apply vaseline. After being burned by phosphorus, you can wash the injured area with 1% silver nitrate solution, 5% copper sulfate solution, or potassium permanganate solution, and then bandage it. Do not rinse with water;
30. When chemical reagents accidentally splash into the eyes, immediately rinse the eyes with flowing water for 15 minutes. If the symptoms do not relieve, continue rinsing for 15 minutes. Immediately apply eye drops after rinsing. In severe cases, send to the hospital for treatment; (III, IV)
31. When a large amount of corrosive chemical reagents or poisons that can be absorbed through the skin come into contact with the skin, immediately rinse the whole body with the emergency shower for 15 minutes. In severe cases, send to the hospital for treatment; (IV)
32. In case of a safety accident involving contact with chemical reagents, the accident record should be filled out in a timely manner.
33. When a small-scale fire occurs during heating the sample or in the experimental process, immediately extinguish the open flame with a wet asbestos cloth or wet rag, and unplug the power supply and turn off the power switch;
34. When a solid of a flammable liquid (mostly organic matter) is on fire, it must not be extinguished with water; (IV)
35. If the burning substance is a flammable gas, such as gas, liquefied petroleum gas, water, acid-base fire extinguishers and foam fire extinguishers are all ineffective. "1211" fire extinguishers and dry powder fire extinguishers should be used. "1211" is a halogenated alkane fire extinguisher, which can form highly toxic phosgene at high temperatures. Pay attention to anti-poisoning when using it. However, they have the characteristics of good insulation, no traces left on the burning object after extinguishing the fire, and no damage to the instrument and equipment, etc., and are suitable for extinguishing fires of precision instruments, precious books and materials and wires, etc.;
36. When the burning substance is a flammable metal, such as potassium, sodium, calcium, magnesium, etc., it is prohibited to use water and acid-base or foam fire extinguishers. In the case that carbon dioxide fire extinguishers, dry powder fire extinguishers and "1211" fire extinguishers are all ineffective, dry sand and 7150 fire extinguishing agent should be used;
37. When a small-scale fire occurs during heating the sample or in the experimental process, immediately extinguish the open flame with a wet asbestos cloth or wet rag, and unplug the power plug and turn off the main power switch and gas valve. When a solid of a flammable liquid (mostly organic matter) is on fire, it must not be watered. In case of a large-scale fire, it should be extinguished immediately with fire-fighting sand, foam fire extinguishers or dry powder fire extinguishers. For extinguishing fires of precision instruments, carbon tetrachloride fire extinguishers should be used. Fires in the laboratory are not suitable for extinguishing with water.
38. Cut off the power supply quickly. If the power supply cannot be cut off, use dry wood strips or insulating rubber gloves and other materials to quickly pull the person who has been electrocuted away from the power supply;
39. Quickly transfer the person who has been electrocuted to a nearby appropriate place, unbutton their clothes to make their whole body stretch;
40. If the person who has been electrocuted is in a state of shock and the heart stops beating or breathing stops, artificial respiration or cardiac massage should be performed immediately. Regardless of whether there are external injuries or not, they should be sent to the hospital for treatment immediately.