Team:XHD-Wuhan-B-China/Safety
Safety
Training
In order to ensure the smooth progress of the experiment and the safety of the students participating in the experiment, we conducted laboratory safety training before we enter the laboratory to work.
1. Strictly abide by the rules and regulations of the laboratory.
2. A preview must be done before the experiment.
3. Take care of public property and save water, electricity, medicines, reagents and raw materials.
4. Check in every day, and the responsibility belongs to the person.
5. Pay attention to safety rules during the experiment.
6. At the end of the experiment, tidy up the experiment table, and do not bring out any items from the laboratory.
7. Responsible for cleaning after the experiment.
Pay attention:
1. Wear lab coats in the laboratory.
2. Don't wear slippers and open-toed sandals.
3. Unclear reagents and instruments should be used after consulting the teacher. Some reagents are poisonous and should not be used at will.
4. Please refer to the following experimental steps to explain the standard use of the instrument.
5. It is forbidden to eat or drink in the laboratory!
Because the microorganisms involved in our experiments have the risk of being released into the environment, when we discard the microorganisms used in the experiment, we cannot directly discard them. They need to be treated with harmless treatment before they can be discharged.
Laboratory protocol
1. Do not play around, smoke, or eat in the laboratory. You must wear laboratory clothes, masks, gloves and other protective equipment when doing experiments.
2. The glassware and reagents used in the experiment should be placed in a reasonable and orderly manner, and the experimental table should be kept clean and tidy. When the experimental work is over or temporarily ended, the utensils and reagents should be returned to their original places.
3. Flammable and explosive reagents must be stored in a cool and ventilated place, and should not be exposed to the sun or close to heat sources.
4. When the skin or clothes are stained with chemical reagents, they should be rinsed immediately with water. If the eyes are sprayed, they should be rinsed immediately with plenty of water. However, if concentrated sulfuric acid drips on the skin, be sure to wipe off the beads with a rag before using it. Rinse with plenty of water.
5. When using glass instruments, check whether there are cracks and whether there are sharp edges or corners to reduce accidents. When using glass instruments, they should be handled with care to prevent damage to the instrument.
6. The waste of small amount of toxic reagents must be flushed into the sewer with a large amount of water, and the waste acid and waste alkali with higher concentration must be neutralized before being discharged into the sewer.
7. The name, label, date of manufacture, grade trademark, factory name, certificate of conformity, etc. of the purchased chemical reagents should be checked one by one. Products with "three noes" and products beyond the warranty period shall not be inspected and stored in the warehouse.
8. Chemical reagents that have been accepted without error should be stored separately according to general chemical reagents, highly toxic substances, flammable and explosive products, strong oxidants, strong corrosives, etc., and should not be piled up and down, and acid and alkali should not be put together.
9. Chemical reagents should be stored in a cool, dry, ventilated place away from direct sunlight. Reagents that require refrigeration should be stored in the refrigerator.
10. The test solution, titration solution, standard solution, etc. must be operated in strict accordance with the relevant standards. After the solution is prepared, an obvious label should be affixed. The content of the label includes the name of the product, the concentration of the preparation, the date of preparation, the preparation person, etc., if necessary, please indicate Validity period and special storage conditions.
11. Alkaline solutions and concentrated salt solutions should not be stored in glass bottles with ground stoppers, so as to prevent the stoppers from being corroded or hard to open after being solidified. Solutions that are easily decomposed when exposed to light should be stored in brown bottles and kept in a dark place.
12. The glass instruments used in the experiment should be kept clean at all times. When they are damaged, they should be placed together to avoid cuts to the staff. The glass instruments should be cleaned in time after use. Do not leave fat, acid, alkali or corrosive substances in the instrument. Or toxic substances.
13. For non-standard mouthed glass instruments with stoppers, such as volumetric flasks, colorimetric tubes, iodine measuring bottles, etc., the stoppers should be tied to the tube with plastic rope or rubber band before washing, so as not to break the stopper or mess with each other.
14. When using a cuvette, only the frosted surface can be used, and the translucent surface cannot be held by hand. When comparing the color, the external water droplets should be absorbed with filter paper, and then wiped with lens paper. Do not use filter paper to wipe the translucent surface with force.
15. Before leaving work, check whether the water and electricity are shut down and make sure that there is no safety concern before leaving.
1. Strictly abide by the rules and regulations of the laboratory.
2. A preview must be done before the experiment.
3. Take care of public property and save water, electricity, medicines, reagents and raw materials.
4. Check in every day, and the responsibility belongs to the person.
5. Pay attention to safety rules during the experiment.
6. At the end of the experiment, tidy up the experiment table, and do not bring out any items from the laboratory.
7. Responsible for cleaning after the experiment.
Pay attention:
1. Wear lab coats in the laboratory.
2. Don't wear slippers and open-toed sandals.
3. Unclear reagents and instruments should be used after consulting the teacher. Some reagents are poisonous and should not be used at will.
4. Please refer to the following experimental steps to explain the standard use of the instrument.
5. It is forbidden to eat or drink in the laboratory!
Because the microorganisms involved in our experiments have the risk of being released into the environment, when we discard the microorganisms used in the experiment, we cannot directly discard them. They need to be treated with harmless treatment before they can be discharged.
Laboratory protocol
1. Do not play around, smoke, or eat in the laboratory. You must wear laboratory clothes, masks, gloves and other protective equipment when doing experiments.
2. The glassware and reagents used in the experiment should be placed in a reasonable and orderly manner, and the experimental table should be kept clean and tidy. When the experimental work is over or temporarily ended, the utensils and reagents should be returned to their original places.
3. Flammable and explosive reagents must be stored in a cool and ventilated place, and should not be exposed to the sun or close to heat sources.
4. When the skin or clothes are stained with chemical reagents, they should be rinsed immediately with water. If the eyes are sprayed, they should be rinsed immediately with plenty of water. However, if concentrated sulfuric acid drips on the skin, be sure to wipe off the beads with a rag before using it. Rinse with plenty of water.
5. When using glass instruments, check whether there are cracks and whether there are sharp edges or corners to reduce accidents. When using glass instruments, they should be handled with care to prevent damage to the instrument.
6. The waste of small amount of toxic reagents must be flushed into the sewer with a large amount of water, and the waste acid and waste alkali with higher concentration must be neutralized before being discharged into the sewer.
7. The name, label, date of manufacture, grade trademark, factory name, certificate of conformity, etc. of the purchased chemical reagents should be checked one by one. Products with "three noes" and products beyond the warranty period shall not be inspected and stored in the warehouse.
8. Chemical reagents that have been accepted without error should be stored separately according to general chemical reagents, highly toxic substances, flammable and explosive products, strong oxidants, strong corrosives, etc., and should not be piled up and down, and acid and alkali should not be put together.
9. Chemical reagents should be stored in a cool, dry, ventilated place away from direct sunlight. Reagents that require refrigeration should be stored in the refrigerator.
10. The test solution, titration solution, standard solution, etc. must be operated in strict accordance with the relevant standards. After the solution is prepared, an obvious label should be affixed. The content of the label includes the name of the product, the concentration of the preparation, the date of preparation, the preparation person, etc., if necessary, please indicate Validity period and special storage conditions.
11. Alkaline solutions and concentrated salt solutions should not be stored in glass bottles with ground stoppers, so as to prevent the stoppers from being corroded or hard to open after being solidified. Solutions that are easily decomposed when exposed to light should be stored in brown bottles and kept in a dark place.
12. The glass instruments used in the experiment should be kept clean at all times. When they are damaged, they should be placed together to avoid cuts to the staff. The glass instruments should be cleaned in time after use. Do not leave fat, acid, alkali or corrosive substances in the instrument. Or toxic substances.
13. For non-standard mouthed glass instruments with stoppers, such as volumetric flasks, colorimetric tubes, iodine measuring bottles, etc., the stoppers should be tied to the tube with plastic rope or rubber band before washing, so as not to break the stopper or mess with each other.
14. When using a cuvette, only the frosted surface can be used, and the translucent surface cannot be held by hand. When comparing the color, the external water droplets should be absorbed with filter paper, and then wiped with lens paper. Do not use filter paper to wipe the translucent surface with force.
15. Before leaving work, check whether the water and electricity are shut down and make sure that there is no safety concern before leaving.
A new protocol for safer DNA electrophoresis
This year, we worked together with HZAU-China-2021 to build a safer protocol for DNA electrophoresis. Many nucleic acid dyes such as Ethidium Bromide (EtBr) and Acridine Orange (Goldview) , which are two DNA dyes commonly used in Chinese laboratories, are notoriously dangerous for users due to their carcinogenicity. Some products are safer, yet too expensive for most start-up labs to consume. We cooperated with HZAU-China-2021 to find a relatively safer and cheaper product, GelRed®, for beginners who are not very familiar with working in the contaminated zone.
After testing it and doing some investigations online, we concluded that the only problem of GelRed® is that it interferes with large DNA stripes and causes them to bend or blur. GelRed® and GelGreen® are based on large, cell membrane-impermeant molecules, which are fundamentally safer than small molecules like EB, AO, and SYBR. Large molecules can “drag” DNA to swim a little bit slower. Short DNA fragments are generally saturated with GelRed®, where long DNA fragments are relatively unsaturated with GelRed® and can be unevenly dragged. This theory can explain why short fragments stained by GelRed® looks so much better than large fragments.
However, some simple changes in experimental procedures can make up for this shortcoming. For example, when running large DNA fragments, we can slightly increase the GelRed® usage to provide saturation. Meanwhile, based on our observation, 50μl PCR products are less “good-looking” than 30μl PCR products, probably due to the different width of the gel poles.
So, our new protocol is:
1. Be really good at designing primers! If your PCR product has only one stripe, no mater how ugly it is, you can always have a successful purification!
2. Use 30μl PCR system.
3. When running large DNA fragments, slightly increase GelRed® usage.
4. Choose short fragments for colony PCR.
5. Choose high-fidelity DNA polymerase over endonuclease when possible.
6. Just because you have GelRed®, doesn't mean you can downgrade the safety procedures of electrophoresis zone in your lab.
After testing it and doing some investigations online, we concluded that the only problem of GelRed® is that it interferes with large DNA stripes and causes them to bend or blur. GelRed® and GelGreen® are based on large, cell membrane-impermeant molecules, which are fundamentally safer than small molecules like EB, AO, and SYBR. Large molecules can “drag” DNA to swim a little bit slower. Short DNA fragments are generally saturated with GelRed®, where long DNA fragments are relatively unsaturated with GelRed® and can be unevenly dragged. This theory can explain why short fragments stained by GelRed® looks so much better than large fragments.
However, some simple changes in experimental procedures can make up for this shortcoming. For example, when running large DNA fragments, we can slightly increase the GelRed® usage to provide saturation. Meanwhile, based on our observation, 50μl PCR products are less “good-looking” than 30μl PCR products, probably due to the different width of the gel poles.
So, our new protocol is:
1. Be really good at designing primers! If your PCR product has only one stripe, no mater how ugly it is, you can always have a successful purification!
2. Use 30μl PCR system.
3. When running large DNA fragments, slightly increase GelRed® usage.
4. Choose short fragments for colony PCR.
5. Choose high-fidelity DNA polymerase over endonuclease when possible.
6. Just because you have GelRed®, doesn't mean you can downgrade the safety procedures of electrophoresis zone in your lab.
