Team:IISc-Bangalore/Safety

Safety

Overview

The safety of the members of the team as well as people who might be unknowingly getting exposed to agents from the lab was given priority over everything else over the duration of our project. Other than the general lab safety rules, the following necessary precautions were taken.

Spatial separation

The iGEM 2018 lab at IISc is divided into two parts.

The first part is exclusively dedicated to molecular biology and biochemistry work and is equipped with two Biosafety level 1 Laminar airflow cabinets. This part of the lab has all the necessary lab safety equipment like an emergency shower, eye washes and fire extinguishers that might be needed in case of an emergency. The second part of the lab is dedicated to phage work and handling mammalian cell lines. This lab is occupied with a Biosafety hood and separate incubators and refrigeration units as well as all the important safety equipment.



The following disposal guidelines were followed for the waste generated during the project. (Excluding phage waste)

  • Liquid waste (cultures, cell pellets etc.): The liquid was treated with 40% sodium hypochlorite and disposed into a separate container for biological waste disposal.
  • Plastic waste: All the plastic waste, including used pipette tips, microfuge tubs etc. were allowed to stand in a closed container containing 40% sodium hypochlorite overnight followed by disposal into a container for plastic waste disposal.
  • Plates: All plates were autoclaved at 121°C for 20 mins to remove the agar as liquid and were washed with detergent and water.

T4 bacteriophage

Use of the T4 bacteriophage presents a critical risk to the rest of the work being carried out in the lab. Even a single virion can completely lyse a batch culture that is being used for protein expression or plasmid preparation. As such, we decided to carry out the work in two separate lab spaces and made sure to segregate both the waste and the containers used for making the media and other buffers.

The following disposal guidelines were followed for disposal of phage waste.

  • Liquid waste (infected cell cultures etc.) : The liquid was treated with 2M NaOH and disposed into a separate container reserved for phage waste.
  • Plastic waste: All the plastic waste, including used pipette tips, microfuge tubs etc. were allowed to stand in a closed container containing 2M NaOH overnight followed by disposal into a container for phage specific plastic waste disposal.
  • Plates: 3mL of 2M NaOH was added to the petri plates and left overnight. This was followed by wash with detergent and water the next day.

mcp-1 chemokine

The mcp-1 protein is a potent chemoattractant for monocytes, a class of cells responsible for phagocytosis of pathogens (after differentiation into macrophages). While our recombinantly produced mcp-1 is the mouse homolog of the human counterpart, it has been shown to have considerable efficacy in recruiting human macrophages too. Considering this, all the work related to mcp-1 was carried out separately from other protein work and proper care was taken to prevent its release into other buffers or the lab space.

Chassis organisms

All the chassis organisms used during the project are Risk group 1 organisms, these being.

  • E. coli DH5α
  • E. coli BL21 (DE3)
  • E. coli BL21
  • E. coli MG1655
  • T4 phage