Trondheim iGEM 2013


The university has a health, safety and environment (HSE) group with set guidelines and rules to regulate the working environment and safety. General information about this can be found at Details on the HSE regulations for the lab work can be found at These are all based on the national biosafety regulations and guidelines set by the Norwegian Biotechnology Advisory Board [3] and the acts set by the government.

Lab Work

The Department of Biotechnology has set local guidelines and rules that apply for the specific labs that we use in our lab work for this project. The labs used are approved for work with genetically modified organisms (GMOs). Prior to the commencement of work in any of these labs, every person is given the essential training. The training covers the statutory regulations and safety provisions, the use of equipment and protective equipment, fire extinguishing, first aid, use of the chemical substances index and safety data sheets, proper waste disposal and reporting. The wetlab part of the NTNU iGEM team has all attended this training and is following the set lab guidelines and rules in their work.

We, of course, follow general lab safety and wear lab coats, disposable gloves and safety glasses when required by the safety guidelines and rules. The materials used in this project are mostly standard kits, restriction enzymes and buffers that are bought and come pre-made from biotechnology companies, and does not require any other safety precautions than the general ones already described above. This also goes for the preparation of solutions like growth medium and gels for gel electrophoresis. When doing gel electrophoresis we use GelGreen instead of ethidium bromide, as it is said not to penetrate cell membranes and thus should not be able to work as a mutagen. However, to be on the safe side, we always wear nitrile gloves when working with GelGreen.

Safety assesment of our project

Our project does not inflict with any of the regulations, guidelines and acts set by the university, institute and government. None of the bacteria and BioBricks used or made in this project raises any serious safety issues for the public health, environment and security. Our activity and use of biological material corresponds to class 1 in the Norwegian regulation for protection against exposure to biological agents and use of GMOs. Class 1 cover use that does not involve any significant risk for human's and animal's health and environment. This corresponds to WHO risk group 1 in the classification of infective microorganisms.

Our aim in one part of this project is to create new combinations of fluorescent proteins (FPs) by manipulating E.coli. FPs is not considered harmful or related to pathogenicity or toxicity. We only worked with nonpathogenic class 1 bacteria; E. coli strain DH5α and ER2566. They do not carry F factor (fertility factor) and are therefore unable to perform conjugation. As the OMVs we are handling are produced by non-pathogenic bacteria and these OMVs has not been reported to have any safety issues, we concluded that they do not provide any pathogenic og toxic hazards.

Plasmids carrying genes for antibiotic resistance are used for selection, and the bacteria carrying these plasmids could in theory be of health and environmental concern if not handled properly. As a golden rule, GMOs should not under any circumstances be brought outside the lab or outside a safe container. To overcome any possible safety issue in relation to the points described above, all biological material and equipment that has been in contact with this biological material are contained in special waste containers within the lab and autoclaved before disposal.

Our aim in the other part of our project is to introduce protein G from S.dysgalactiae subsp. Equisimilis into E.coli Outer Membraine Vesicles (OMVs). S.dysgalactiae subsp. equisimilis is an opportunistic pathogen that causes mastitis and polyarthritis in animals, and in rare cases, it can cause meningitis in humans. Before starting working with protein G we carefully investigated the risk of the protein as it comes from a pathogenic bacterium. We did research in the literature, which considered protein G to be a part of S.dysgalactiae subsp. equisimilis’ pathogenesis but not a pathogenic determinant[1]. We consulted an expert on protein G, Inga-Maria Frick from Lund University, who considered our experiments to be safe. I avoid work with pathogenic bacteria we asked Kjersti Wiik Larsen at St. Olav Hospital to prepare the S.dysgalactiae subsp. Equisimilis genome samples that we used as a templates in PCR reactions.

If any of our organisms where to be released to an environment outside the lab, either by design or accident, the biggest threat in theory would be the transferring of genetic material and conferment of antibiotic resistance to other microorganisms in natural habitats and microorganisms related to human, animal or plant diseases. Antibiotic resistance is of huge concern when it comes to the treatment of infectious diseases and the risk of this happening should always be taken seriously and appropriate safety steps should be made. The steps described in this text, together with the set guidelines and rules, are considered sufficient to prevent this.

Risk Assesment

The following risk assessments for the lab procedures we are using have been made by the institute’s HSE group. The grading system for personal and environmental risk assesments ranges from A to E, where A is no/very low risk, B is low risk, C is moderate risk, D is high risk and E is very high risk:

Activity Safety Procedures Personal risk Env. risk Comment
Agarose gel electrophoresis, (GelGreen) Nitrile gloves, protective eyewear (with UV filter), face shield when needed. GelGreen used for staining GelGreen: Unknown, UV: B A GelGreen is said not to penetrate cell membranes, and thus should not act as mutagen even if it is DNA-binding. Gloves also minimize the risk for exposure. UV damages on unprotected skins/eyes if instructions not followed.
Antibiotic-stock solution, make and use of Gloves, handle powder only inside fume hood A B May cause allergic reactions if instructions not followed. May causlead to multi-resistant bacteria if not disposed correctly.
Autoclave Thermoresistant gloves, Eyeprotection lab, Instructions posted for not opening autoclave too early and for not overfilling bottles or closing their lids completely. C A Rapid pressure fall due to opening the autoclave to soon may cause hot liquid burns on eye or skin. Will not happen if instructions are followed.
Bacteria class 1 and recombinant bacteria Autoclave accessible. Inactivation of genmanipulated bacteria in contaminated material and waste. Labcoat mandatory. Lab bench surfaces resistant to water, acid, alkali, solvents, disinfective agants, decontaminating agents and easy to clean. Transport between labs only in closed containers. Good microbiological practice. A A Risk include Release of GMO to environment, bacterial infections. Low because DH5 alpha are crippled.
Use of open flames – (e.g. sterilization with bunsen burners) Bunsen burner must not be left burning B A Risks include skin burns and fire. New rules on handling installed.
General lab work Safety rules according to risk assessment - - No injuries requiring more than simple first aid in these laboratories for the past 5 years, the present rutines seem sufficient
DNA isolation and purification Use gloves and follow the safety tips based on the kit manual - - PB buffers should not be mixed with the bleach.
Preparation of medias for growing bacteria According to MSDSs of relevant chemicals - - Depends on the chemicals
PCR Fume hood when DMSO is added A A PCR-machine should be in ventilation hood when DMSO is added, use lab coat and gloves
SDS PAGE gloves, lab coat, ventilation hood when adding dye - - Put the SDS PAGE chamber under the ventilation hood
Centrifugation (Sorvall + table) Accurate balancing, accurate attachment of rotor, not exceed maximal G-forces fore each type of tube C A Danger is damage caused by loose rotor
Ventilation hoods, use of Opening minimized when not in use, correct settings when in used for protection C A Exposure to hazardous chemicals due to insufficient airflow (effects on local hood or other hoods). Can be prevented by maintaining sufficient airflow

Use this page to answer the questions on the safety page.

Safety forms were approved on September 24, 2013 by Evan Appleton.