Every member in GreatBay_SZ has learned basic lab access and rules. Besides, we put personal biosecurity at high priority when working through our experiments. Working together with team GreatBay_SCIE and team Genas_China, we arrange members to be in charge of the daily cleaning of lab equipment and safety check. Our team set up detail rules towards equipment usages, waste management and specific procedures and protocols that help us maintain lab safety. Under the instruction of our professor, we set up few penalty rules for members who did not follow basic lab safety, for the overall purpose of increasing the efficiency in lab safety management.
Safety in Experiments
The three Ecoli strains our team used to design are in biosafety level 1, including Ecoli DH5a used for basis DNA design, Ecoli BL21(DE3) use for synthesizing spider silk protein, and Ecoli MG1655 for synthesizing natural pigments. All three strains are the most common used chassis in the iGEM white list, and all of our members have been trained with basic rules to work with Ecoli. We pay high attention to avoid any of our organisms escape from the lab, and even if organisms escape they cannot survive long without specific conditions.
Protein Purification From Cells
The spider silk our team acquired has gone through his-tag purification, and in addition we use lysosome to break down all bacteria remains and filtrated all none-protein substance. Some sample of protein will be lyophilized and stored for later usage.
We extract deoxyviolacein dye by lysis the bacteria with NaOH and neutralize with acetic acid, we have centrifuged the mixture and filtrated all the remaining impurities. The next step we uses ethanol to mix with our dye extract, to make sure all bacteria are dead and no recombinant DNA will remain in our dye. Some sample of deoxyviolacein will be lyophilized and stored for later usage.
The spinning machine didn’t contact with any pathogen, chemicals or flammable material. The only two materials that required when spinning the protein are Tris buffer and isopropanol, which are harmless to human.
Possible Risk and our solution:
- During the process of fermentation, bacteria may be inhaled that might cause potential vomit and diarrhea. Our team make sure that all cell cultures, if put in open lab space, will be sealed with no exposure for bacteria to escape from the flask or plate.
- During the SDS electrophoresis of protein gel, physical contact with acrylamide might lead to chemical poisoning. - Our team makes sure that every member before contacting with acrylamide need to wear a mask, in addition to lab-gown and gloves.
- During the process of spinning spider silk, syringe needles will be used and it makes cause minor injuries. Our team make sure that the spinning machine will only be used when a supervisor is around.
Safety in Human Practices
Since this year we extract deoxyviolacein and indigo dye in our lab, we invite a few people to come and try to design and use our dye. All the sample of pigments and dyed textiles/works were being kept inside the lab with a carefully constructed sterilized measurement. Our team makes sure that none of our organisms or any material that might likely contact with our bacteria will be released out of the lab.
For any visitors that come and into our lab, we taught them basic lab rules, and giving them safety protection tools such as lab-gowns and gloves.