Prologue

As responsible researchers, it is necessary for us to select appropriate organisms for experimentation (which includes avoiding the use of organisms in risk groups 3 and 4), comply with the non-release policy, conduct experiments in appropriate laboratories, and take proper personal protection in the experiments. The followings are NCHU_Taichung’s safety-related information:


Biosecurity

Our project was divided into 2 parts:

  1. Biosynthesis of PQQ
  2. HaCaT cell wound healing assay
    • We transferred the plasmid with PQQ (Pyrroloquinoline quinone) biosynthetic gene domain into Bacillus subtilis var. natto (BS) and let the bacteria express. Then proper extraction methods were used to conduct a quantitative analysis of PQQ, which is a high-value polyquinone with multiple bio-function in organisms.
    • HaCaT cells were used to mimic skin wounds under natto extract containing PQQ and to observe the characteristics it represented. The bacteria would be killed in the extraction step and only remain in the product in the medium for further investigation, so none of our parts could be hazardous to the users. Additionally, the chassis used in the above steps are 1. Escherichia coli JM109 and 2. Bacillus subtilis. Bacillus subtilis is listed on the iGEM White List.

Chemical Control

Among the chemicals used in the experiments, including acetic acid, sodium hydroxide, phenol: chloroform, trizol, ethanol absolute, trypan blue, methanol, hexane, benzalkonium chloride, and pentanol are used and discarded according to the regulations and guidelines of NCHU’s Safety Center (full name: Center for Environmental Protection & Occupational Safety and Health). The experimental operations were all carried out in a fume hood, equipped with personal safety protection (see Protective equipment for details).

Protective Equipment

It is necessary to be equipped with qualified personal safety protection equipment (PPE) during the experiment and in qualified environments. We have gloves, goggles, and lab coats during the experiments.


Disposal

  1. Biological waste is sterilized in an autoclave at 121°C for 20 minutes and discarded according to the rules of the NCHU Safety Center.
  2. The disposal of chemicals is managed by university officials who transmits the waste to a licensed third party.

Safety Training

All the wet lab members of NCHU_Taichung would perform the experiments after taking proper laboratory training, including the use of a pipet, the safe way to execute microorganisms, the use of emergency instruments, and the right process while using high-speed centrifugation.