Team:Uppsala/Safety

When working with class 1 bacteria it is important to consider the potential harm it can cause, both towards the students and the environment. That why we had several precautions to make sure the risk was as limited as possible. The lab worked in is classified as biosafety level 1. This allowed us to work with non-pathogenic bacteria such as Escherichia coli TOP10 and BL21(DE3)*. To limit the spread of bacteria in the lab, 70% ethanol was frequently used on benches and other surfaces as well as the lab crews hands to disinfect. This way the bacteria is killed off straight away and its risk of spreading both in the environment and in us is reduced. All liquid bacterial cultures and LB/SOB that may have been contaminated were treated with “Jodopax”, which is an iodine rich substance that kill bacteria. The Jodopax was left in the bacterial solution for at least 1h before disposing of it in the sink. All glass pipettes used were put into buckets with “Virkon” solution to ensure sterilization. They were left in the buckets for at least one day.

When working with phages we kept to a single fume hood. We had designated equipment for working with the viruses, and we wiped surfaces with chloroform.

Hazardous chemicals

Acrylamide:
Acrylamide is used in SDS-PAGE. The substance is toxic which can cause skin and eye irritation and can cause after long exposure organ damage. To protect us when using the substance thick nitrile gloves, safety glasses and lab coats were used. When handling the substance the one working with it was always extra cautious and disposed anything that might have come in contact with the substance in a special hazardous waste container.

Antibiotics:
Antibiotic is classified as a drug according to swedish law and is currently increasing in resistance in the environment. When handling the substance gloves, lab coats and glasses were worn and material that was used in contact with the substance were sorted into contagious waste boxes. Small liquid cultures including antibiotics were treated with “Jodopax” to destroy the chemical.

Liquid nitrogen:
Liquid nitrogen was used to snap freeze when making our competent cells. When liquid nitrogen is very cold it has a boiling temperature of −196 °C which means it needs to be handled with care. That’s why a special thermos was used to make sure the gaseous nitrogen could escape to keep the pressure low. When liquid nitrogen was used it was handled with care and safety goggles, lab coat and insulating gloves were worn at all times.

Nickel Sepharose Fast Flow:
The substance contains nickel, which can cause allergic reactions when come in contact with. The nickel ions are also flammable. The substance was kept away from fire and heat at all times. The substance was handled with care and gloves were worn at all times. Contaminated liquid was disposed contagious waste boxes as nickel is dangerous for the environment.

PDMS:
PDMS or Polydimethylsiloxane is a silicon-based organic polymer. The product is non-toxic below 180 ℃. Higher temperatures may release hazardous gases. However it was never used at such high temperatures. The product can cause eye irritation so glasses, gloves and lab coats were worn at all times.

Chloroform:
Chloroform was used to clean PDMS as well as for cleaning where P1 phage was used. The substance evaporates at room temperature and is dangerous. That why all work with the substance was done in the fume hood with clearly marked bottles. Nitrile gloves, glasses and laboratory coat were worn.