Team:SZTA RMG Szeged/Safety
Safety
Contents
I. Proposed product safety
1. Background considerations
As we intentionally use random RNAs and DNAs which were checked not to interfere with the human genome, we expect that these initial experiments won't mean any serious hazard to any person or to the environment. As our detection kit doesn't contain any truly or potentially living organism, it cannot cause any biohazard. We will also use polymerase, restriction enzymes and the buffers they require and the SYBR Green II dye which is characterized as "not hazardous" according to the supplier's MSDS. After the experiments, the samples and the reaction mixtures are going to be fertilized and destroyed according to the Hungarian.
2. Proposed way of recycling the main component
Research suggests that over 95 degrees Celsius the magnetic beads are no longer able to immobilize DNA, hence allowing us to use the beads more than one time to detect RNA. This significantly decreases the price of the method, given that the magnetic beads are the most expensive component of the detection, and it also makes the detection eco-friendlier.
Recycling the magnetic beads protocol:
-After the previous detection is over, do not discard the magnetic beads, instead resuspend them in 1mL 1X Binding & Washing Buffer (for more information see our Protocol)
- Incubate the solution on 95 degrees Celsius for 30 minutes
- Put the test tube with the solution onto the magnet
- Discard the supernatant
- The magnetic beads are no longer binded to ssDNA, they are ready to be used for further detection, by repeating the protocol from the beginning
II. Lab Safety Measures
1.Safety Education
At the beginning of the lab work the team has participated in a thorough biosafety education held by our mentors. The main idea of this course was to clarify that working in a molecular biology laboratory has many other important aspects beyond the scientific research work. We went through all the possible personal, public and environmental risks and possible coping strategies, as well. We have learned how to use the fire extinguisher, how to use chemically and/or biologically unsafe materials and how to handle safely all the manually or electronically controlled equipment.
2.General approach
We worked in an organized laboratory, in an RNase-free environment. Therefore, washing with RNaseZap was an integral part of starting work. Prior to laboratory work, everything was treated with an RNase inhibitor to ensure an optimal, RNase-free environment. Laboratory coats, rubber gloves, and masks (FFP3) were an indispensable part of everyday laboratory equipment. They were properly discarded after use, which is a crucial part of laboratory safety. Furthermore, we paid paramount attention to keeping the solutions RNase free, by opening the test tubes for the shortest time possible. We also followed instructions regarding the storage of the components used.
3.Waste management
All the DNA and RNA samples and other disposable materials (pipette tips, Eppendorf- and PCR-tubes) connecting with them were dropped into a container, in which strong acidic environment damaged them. After this step, all our waste is going to be transported by an officially approved firm which will continue the safety procedures with our used materials adhering to the Hungarian regulations.
Nevertheless, we always pay attention to the following circumstances when we use it:
- we use all personal safety equipment (personal lab coat, safety glasses, disposable gloves). We always work with it in a sterile box to avoid the inhalation of any cells or spores. Moreover as it may be harmful to any small skin injuries by causing local inflammations, we constantly fertilize our hands.
- we fertilize all desks and benches just after each lab sessions, to avoid the survival of any cells/spores
- even though these strains can't survive in the environment (or at least it has a very low probability), all cultures are handled carefully according to the local lab regulations. After using the cultures, all affected disposable tools and liquids (centrifuge tubes, pipet tips, reaction mixtures, etc.) are sterilized by an autoclave and collected to sealed boxes which are transported by a decontamination company each month. All non-disposable glass tools are bleached for 24 hours and are autoclaved according to the lab regulations.
4.Chemical safety
- Our kit uses exclusively not harmful reagents - we should pay attention in the user's manual on the proper use with a clear protocol.
- As it may happen that the kit will be used with potentially harmful samples (soil / wastewater / human samples / etc. - in which the kit can detect the target ssRNA) we should strongly suggest to the users to be aware of the antiseptic techniques, too.
- After the detection process the reaction mixtures and samples should be handled according to the local regulations - the main aim is to destroy the potentially hazardous samples appropriately.
- After the detection process the reaction mixtures and samples should be handled according to the local regulations - the main aim is to destroy the potentially hazardous samples appropriately.
5.Freight of the kit
- As the kit contains temperature sensitive enzymes / nucleic acids it should be shipped in dry ice - therefore we should ask the users to pay attention when the package is being opened.
- As the kit itself doesn't contain harmful materials, it doesn't require any havaria management plan.
Ethical risk:
- The users should be aware of the kit's detection accuracy in the terms of sensitivity and specificity which should be communicated clearly in the user's manual.
6.Electric safety
- As the device is planned to be operated at low voltage, normal batteries should be available. As the batteries usually may be harmful for the environment we should think about the usage of rechargeable batteries and solar panels for charging.
We have used microorganisms belonging only into BSL1 level. Concerning genetically modified organisms, we had to fulfil the requirements of the rather strict Hungarian State regulations. (You can read more on these regulations here: http://gmo.kormany.hu/hungarian-legislation)
