Team:ZJUT-China/Safety
Human Practices
Safety
- 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. Security issues have always been our focus. Before we started the project, 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. So ，our team designed and operated the experiment strictly according to the laboratory safety standard, which includes formulating strict laboratory safety criteria and，specially，the safety of bacteria used in our experiment.
Project design safety
- Our designs this year include indicator group, formaldehyde degradation group and photo cracking group, all of which meet safety standards.
- Non-pathogenic strains
For the safety of our experiments, we select E. coli BL21,DH5α,E.coli MG1655 as our host organism in our project. These chassis microorganisms are commonly utilized in laboratories and have not possessed any characteristics that can endanger the human body and the security of environment. Thus, our project is safe from the perspective of strains we used.
- Harmless plasmids
The plasmids we use in our project (pGLO-Cas9, pTargetF-p15A, dusk-Cas9-pUC57, dusk-eGFP-pUC57, pGLO-lacI, pGLO-cI,pGLO-lysis for E. coli) are almost harmless for humans. Consequently，plasmids lead to no safety issues.
- Innocuous gene parts
The gene we utilized in our design are all meet the safety requirements. During the time of designing, we ensure that none of the parts assembled into our genetic engineering machine is able to take a toll on health in existent researching files. As a result, we confirm that all parts wouldn’t act as virulence factors.
- Secure genetic circuits
In order to further improve the security of project, a module for cell lysis is constructed to disrupt the cell after eliminating the ARG. We use lysis gene as the functional element, the cells wall will be split once the inducible promoters are induced .Besides, in our project ,we choosed the GFP and eGFP as reporter genes which can be detected directly without adding any harmful reagents.
Lab Safety
- A lab manager (Dr. Zhang Yinjun) is responsible for the safety and security of iGEM lab. He is responsible for the safety and security of biology labs at our institution, Institutional biosafety officers, supervised us, gave us safety training courses in the laboratory, and set up a special management equipment manager, special laboratory equipment (such as high voltage Bacteria pots, high-pressure cylinders and other pressure vessels) will perform inspections, annual inspections, etc., and designate special personnel to hold certificates. The post-employed personnel shall undergo training as required, and after passing the examination, they shall be allowed to take posts with qualified posts, and it is forbidden to operate without certificates. Laboratories need to establish special equipment operation files to ensure safety experiments. These experts have a long history of working with an organism or part that might pose a risk, they are familiar with the experimental procedures and practices we will do.
- Wash your hands with disinfectant soap when you arrive at the lab and again before you leave.
- Absolutely no food, drinks, chewing gum, or smoking is allowed in the laboratory. Do not put anything in your mouth such as pencils, pens, labels, or fingers. Do not store food in areas where microorganisms are stored.
- Purchase a lab coat and safety glasses, bring them to class, and use them. Alternatively, a long sleeved shirt that buttons or snaps closed is acceptable protective clothing. This garment must cover your arms and be able to be removed without pulling it over your head. Leave protective clothing in the lab and do not wear it to other non-lab areas.
- Avoid loose fitting items of clothing. Wear appropriate shoes (sandals are not allowed) in the laboratory.
- Keep your workspace free of all unnecessary materials. Backpacks, purses, and coats should be placed in the cubbyholes by the front door of the lab. Place needed items on the floor near your feet, but not in the aisle.
- Disinfect work areas before and after use with 70% ethanol or fresh 10% bleach. Laboratory equipment and work surfaces should be decontaminated with an appropriate disinfectant on a routine basis, and especially after spills, splashes, or other contamination.
- Label everything clearly.
- Replace caps on reagents, solution bottles, and bacterial cultures. Do not open Petri dishes in the lab unless absolutely necessary.
- Inoculating loops and needles should be flame sterilized in a Bunsen burner before you lay them down.
- Turn off Bunsen burners when not is use. Long hair must be restrained if Bunsen burners are in use.
- When you flame sterilize with alcohol, be sure that you do not have any papers under you.
- Treat all microorganisms as potential pathogens. Use appropriate care and do not take cultures out of the laboratory.
- Wear disposable gloves when working with potentially infectious microbes or samples (e.g., sewage). If you are working with a sample that may contain a pathogen, then be extremely careful to use good bacteriological technique.
- Sterilize equipment and materials.
- Never pipette by mouth. Use a pipetting aid or adjustable volume pipettors. [In the distant past, some lab personnel were taught to mouth pipette. This practice has been known to result in many laboratory-acquired infections. With the availability of mechanical pipetting devices, mouth pipetting is strictly prohibited.
- Consider everything a biohazard. Do not pour anything down the sink. Autoclave liquids and broth cultures to sterilize them before discarding.
- Dispose of all solid waste material in a biohazard bag and autoclave it before discarding in the regular trash.
- Familiarize yourself with the location of safety equipment in the lab (e.g., eye-wash station, shower, sinks, fire extinguisher, biological safety cabinet, first aid kit, emergency gas valve).
- Dispose of broken glass in the broken glass container.
- Dispose of razor blades, syringe needles, and sharp metal objects in the “sharps” container.
- Report spills and accidents immediately to your instructor. Clean small spills with care (see instructions below). Seek help for large spills.
- Report all injuries or accidents immediately to the instructor, no matter how small they seem.
Dispose of items in the special receptacles as indicated below. If you have a question regarding the proper disposal of an item, please ask your course instructor or TA.
Material Method of Disposal Agar slants with biological materiral Place tube upright in indicated text tube rack,but
place caps in basket as indicatedBiological liquids (not in text tubes) Leave in container with closed cap and/or follow
specific instructions given by the TA or course instructorBiological liquids in test tubes Place tube upright in indicated test tube rack with caps left on the tube Broken glass (contaminated) Sharps container Broken Glass (not contaminated) Broken glass container Cotton Swabs (contaminated) Benchtop disinfectant/discard can Needles, glass slides, syringes, pipettes, other types of sharps Sharps container Noncontaminated paper Regular trash Petri dishes and contaminated solids (other than pipettes or swabs Biohazard “orange/red bag” container Transfer pipettes (contaminated) Benchtop disinfectant/discard can
In general, noncontaminated items that pose no threat as a sharp object capable of causing injury can be disposed by placing them in the regular trash. All sharps, contaminated or not, should be discarded into the sharps container. All other contaminated materials have specific locations in which they are discarded.
Overview：
（Dr. Zhang Yinjun）
Microbiology Lab Practices and Safety Rules
