Team:NIT Warangal/Safety

iGEM NIT_Warangal

Safety

Considering the limited access to the laboratory due to the pandemic situation, most of our project work was limited to in-silico tools. Rest of the work was performed in our institute which is equipped with the state of the art laboratory facilities. All experiments were performed according to Good Laboratory Principles (GLP) and procedures put-forth according to the biosafety committee of the institute. Disposal of bio-waste was conducted as per the advice of the biosafety committee.

Laboratory Safety

Our group worked under the direct supervision of Dr. Ashish A Prabhu, the Primary PI of the group. He specializes in metabolic engineering of Pichia pastoris, E. coli, and other strains. All our lab members went through training and guidance under our PI. We followed the standards of institutional biosafety committees framed by the Department of Biotechnology, Government of India, when working with toxins like Gossypol and while handling recombinant strains.

General Lab Safety Measurements

Do not drink or eat in the laboratory.
If you are new to the lab, it is necessary to work under the supervision of your professor or the lab attendant.
Always wear protective gear such as an apron, gloves, and headgear.
Waste disposal should be done properly. It should be well labelled as Bio-hazardous if it is so.
Always take the required amount of chemicals, if it is in access, do not put it back again in the container.
Know where the safety equipment is located such as a safety shower, fire extinguisher, and first aid kit.
Do not mix unknown chemicals.
Be aware of what to do in case of emergency and lab accidents.
Do not leave ongoing experiments unattended.

Project Safety

Our approach is to develop an enzymatic method for efficient degradation of the gossypol without altering other nutrient factors in cottonseed meal (CSM). Before starting the experiments, we followed all the precautionary and safety guidelines as mentioned in iGEM Safety Guidelines and institute biosafety guidelines. We used crude glycerol in nutrient medium for growth of recombinant plasmid. Crude glycerol causes irritation when it comes in contact with the eye and skin. It also causes inhalation problems when used with high concentrations^[1]. We used protective goggles, hand gloves, shoes and an apron to handle crude glycerol cautiously. All the lab instruments were sterilized properly before use. We used Pichia pastoris in our experiments for the production of laccase enzyme to degrade gossypol. Pichia pastoris is neither pathogenic nor toxigenic and is generally regarded as safe. It is also not associated with illnesses to humans and animals. Strains of E. coli DH5 were handled with proper precautions even-though they are non-pathogenic and also due to its ease in cloning experiments. Methanol was used to induce the protein. Repeated exposure of methanol causes chronic poisoning and can be absorbed into the body by inhalation, ingestion, eye contact and skin contact^[2]. So extra care was taken to handle methanol while performing the experiments. After the experiments, the laboratory benches were cleaned with ethanol and reagents. Bio wastes generated were collected and disposed properly according to institute biosafety rules.

References:

[1] Cargill glycerin crude material safety data sheet
[2] https://www.cdc.gov/niosh/ershdb/emergencyresponsecard_29750029.html