Team:Potsdam/Safety
Practical scientific work is crucial, especially for future scientists like us. We like to keep focusing on what science is able to achieve, but then again, risks and dangers may never be out of sight. Science holds a lot of potential to help and improve, but also to damage and destroy. It takes a lot of time, preparation and precaution to avoid situations where the health of people or the environment are endangered.
Thankfully, during our time working in the lab, no hazardous situations occurred, even though most of us were relatively inexperienced in carrying out an experiment by ourselves. This is partially because everyone was taking extra care on what they were doing (and also on what others were doing) and because everyone was required to be well prepared to start working in the lab.
Preparation
Unexpected situations lead to danger, that is why we implemented a seven month preparation time. During this time the topic in general and all experiments were planned in detail. In regular seminars and emails our advisors and instructors were informed about progress and any questions that emerged.
Due to the low practical experience of most of us a project had been worked out that did not contain dangers coming from strains or chemicals.
Only S1 organisms (according to the GenTSV and the GenTG, eng. genetic engineering safety ordinance and genetic engineering law) were used and the practical work took place in an S1 lab. The restrictions say that S1 organisms contain only minimal potential for danger to people and the environment. The strains we used were:
- • E. coli : DH5 α
- • E. coli : JM109
- • S. cerevisiae : YPH500
The plasmids used were:
| • pSB1C3, pSB1A3, pSB1K3 | high copy plasmids carrying the antibiotic resistances to Ampicillin, Kanamycin and Chloramphenicol |
|---|
| • pYES2/CT | Commonly used Yeast expression vector with Uracil auxotrophy |
|---|---|
| • pSB4A5 | low copy BioBrick standard vector with ampicillin resistance |
For the experiments in yeast the backbone pYES2/CT is used. It contains a galactose inducible promotor. Without galactose none of the inserted DNA can be expressed.
The final product in both E. coli and yeast is the hormone Auxin. Auxin is naturally expressed in plants and in small amounts in yeast, where it triggers growth and differentiation of cells. It is neither harmful to people nor the environment if accidentally released.
General Guidelines
In Germany there are many restrictions concerning scientific work with genetically modified organisms (GMOs). In the GenTSV and the GenTG (eng. genetic engineering safety ordinance and genetic engineering law) regulate what is generally forbidden in Germany/in the European Union (EU) and which safety precautions have to be observed. GMOs are defined as “genetically modified organisms, whose genetic material has been altered by molecular biology methods, in a way that would naturally not be possible by breeding or recombination”[1]. We also paid attention to the safety indications given by the iGEM foundation.
Their safety sheets made us aware that the used DNA binding protein dCas9 requires special treatment.
In addition to these guidelines there are regulations in our lab that are set by the university to protect the experimenter and other people in the building. A short summary of those rules, that were explained during an obligatory safety instruction, may be found under “Lab safety”.
In our lab there is a special disposure system that distinguishes between normal waste (paper, packaging, uncontaminated plastic waste), biohazardous waste (might have been in contact with GMOs), chemical waste (e.g. EtBr) and glass. Every kind of waste is treated differently. This way, neither the environment, nor other lab technicians or cleaning staff risk their health while working in the lab.
To give an overview to the university which GMO we are currently working on there are lists, where every new GMO has to be listed. Those lists are also sent to the responsible institution in Brandenburg, GER. They also had an eye on what we were actually doing.
Lab Safety
The best preparations and precautions can never replace paying attention to safety while working. These rules apply to everyone working in a lab of the university of Potsdam.
• Do not eat/drink inside the lab.
• Do not use make up.
• Do not pipette by mouth.
• Wear a lab coat, long pants and closed shoes.
• Keep chairs, bags, cartons etc. out of the evacuation routes.
• Keep windows and doors closed.
• Do not leave potentially dangerous reactions unattended.
• Stay with the used electronic device until it has reached its final speed, temperature, etc.
• Use nitrile gloves, heat or frost protection gloves when needed.
• Do not touch anything with nitrile gloves if not labelled differently.
In order to prevent us from being alone if a problematic situation occurs we were not allowed to be the only workers in the building. This also counted for late shifts and weekends. Among us we had the arrangement to never let anyone be on his own in the lab, especially when we did not share our lab with others anymore.
Our contacts gave us extra instructions for specific devices (e.g. gel electrophoresis with EtBr, gel extraction on UV light). The EtBr especially, that was used to tag our DNA in the gel, had to be handled with care since it is suspected to be a mutagen. It was only allowed to be treated in delimited areas, with labelled equipment and, of course, with gloves. Electronic devices were never to be touched with gloves.
Laptops and phones were mostly kept out of the lab. For seminars and planning we used seminar rooms that are not included in the S1 area. Lab coats had to be left in the S1 area and were only allowed to be taken out after autoclaving. We are quite a large group. Therefore, communication is everything. Keeping everyone up to date made sure no one got confused with chemicals or organisms.
Big thanks to everyone who supported us! You helped us getting our project done safely.
[1] Safety Data Sheet for NEB 5-alpha Competent E.coli (High Efficiency), New England BioLabs, 27-Jun-2016
Safety data Sheet for Bacterial Strain JM109 , Promega Corporation, 02.06.2016
Safety data sheet for (among others) YPH 500 Yeast Strain, Agilent Technologies Manufacturing GmbH & Co. KG, 30/03/2017
[2] https://2017.igem.org/Safety/White_List, 21.09.2017
[3] § 3 (3) s. 1 GenTG as amended on 2017-07-17
Main sponsors:
