Team:TU Eindhoven/Safety/Biosafety

Biosafety

Lab safety

One of the most important things when working in the lab is the safety of the team members and the other persons working in the lab. Therefore it is important to follow the rules of the laboratory.

Because we work with genetically modified organism (GMO) material or with cells in the lab we cannot simply enter the laboratory. So the team needed a safety training first. In this safety training we had a lab tour in which we discussed topics like the safety equipment, personal equipment, safety measures, waste disposal and biosafety rules or guidelines concerns working with GMO’s. After the training we received our permission to enter the lab.

In Europe there are some biosafety regulations, which we also have to follow in the lab. These can be found here. At the Eindhoven University of Technology there is one main person responsible for the biological safety. Her name is Moniek de Liefde – van Beest. She is the biosafety officer of the TU Eindhoven. There are also principal investigators (PI). The PIs are responsible for the biosafety and lab safety at their appointed laboratories.

Some of the biosafety guidelines for our laboratory are as following: when working with GMOs, it is important to be careful, work clean and hygienically, know how to work with GMOs and know the safe microbiological techniques (SMT). SMT ground rules and other biosafety rules at the lab are:

1. Always work according to the rules.
2. Keep doors and windows closed.
3. Gather everything and place it orderly.
4. Keep everything clean and tidy, make sure enough disinfectant is present.
5. Always wear a marked and closed lab coat, which is not permitted outside the lab. The lab coats have to be sterilized by autoclaving before sending it to the laundry.
6. Don’t wear any watches or jewelry, or keep them covered by gloves or lab coat. Bags, laptops etc. are not allowed and can be stored outside the lab in lockers.
7. Avoid any contact between your hands and your face. Don’t eat, drink or smoke inside the laboratory. Storing food or drinks is also not allowed.
8. Avoid formation of aerosols. Mix and centrifuge in closed tubes. The use of a needle is allowed only when no other method is available.
9. Pipetting with the mouth is not allowed; use the available equipment (pipette boy or pipette bulb).
10. Always disinfect your working space at the start and at the end of your activities.
11. After a contamination of your working space (e.g. when you spill any biological material) disinfect the working space.
12. After working with biological agents and when leaving the laboratory, always wash your hands with water and soap.
13. All re-usable materials that were in contact with biological materials have to be sterilized before being washed or discarded.
14. Solid biological material is gathered in the red biohazard bags, transferred in blue containers and disposed of as special garbage. Liquid biological waste has to be autoclaved before it can be discarded.
15. All accidents and spills, and all activities with GMO’s or human material have to be reported in the appropriate logbook.

There are also rules concerning waste disposal: all solid biological waste (ML-I and ML-II) has to be collected in red biohazard bags and transferred to the leak-free blue hospital containers. After closing, labeling and disinfecting the exterior of the container with 70% ethanol it can be disposed of as special waste. Solid waste contaminated with phages can also be collected in the blue hospital containers. Liquid biological waste has to be autoclaved (20 min. at 121 degree Celsius) and can be discarded in the sewer afterwards. Liquid waste contaminated with phages has to be disinfected in a bottle with concentrated chlorine at least overnight. Glass and instruments which have been in contact with biological agents must also be autoclaved and can be cleaned in the dishwasher, subsequently. Glass and instruments which have been in contact with phages must be disinfected overnight with a chlorine solution, and then washed with water, cleaned in the dishwasher and autoclaved. Chemical waste has to be collected in the chemical trashcans in the lab, transferred in the ‘wisseldrum’ near the autoclave and disposed of when it is full.

Biosafety of the Organisms and Parts

The organisms that we used during our project are E. coli (K-12) (Nova Blue), E.coli (B) Bl21 (DE3) and E. coli (K-12) (XL10-Gold). The E. coli K12 strain is a so called ‘lab strain’. This strain cannot survive in the human digestive system and is not able to produce toxins. The Risk group of this strain is 1, and all it derives are also risk group 1 like the used NovaBlue and XL10-Gold strains [1]. The E. coli B and its derivatives, like the used BL21 (DE3) strain, are classified as risk group 1 [4].


The parts that we used are BBa_K1492000, BBa_K1492001 and BBa_K1492002 and are listed in Table 1. For more information check our Biobrick pages (COMPx, COMPy and tRNA Synthetase) and our Safety spreadsheet.