No running, sudden moves within the lab.
No eating within the premises of the lab.
All experiment waste must be thrown in a special bin then put through a machine for sterilization.
All pipettes must be sanitized after use.
All equipment use must follow the using manual.
Performing experiments with a pipette while wearing a lab coat and gloves.
To enter the laboratory, all personnel must wash hand, as well as wear full lab suits and shoes.

Photos of the LAB:

The BIOLAB of Mingdao High School.

Eating and Drinking are not Allowed in the Lab.

The Laminar Flow Clean Bench in the BIOLAB, MDHS.

Warning Signs are Posted in the LAB.

The Emergency Eyewash and Autoclave in the BIOLAB, MDHS.

Our Goal

The high concentration of carbon dioxide in an enclosed space makes people feel sleepy and even dizzy. While driving, drivers may feel asleep, causing accidents to happen. To solve this problem, we plan to use the high photosynthesis efficiency of cyanobacteria to reduce the concentration of CO2. But the solubility of CO2 in water is not high, so we modified Bacillus spp. to produce carbonic anhydrase (CA) from human, which can increase the solubility of CO2 in water. Furthermore, to extend the application, we produce CYP2E1 from rabbits, which is used in xenobiotic metabolism, to eliminate benzene and chloroform. In sum, we built up an efficient microalgae purification system to improve indoor air quality using natural enzymes such as carbonic anhydrase and xenobiotic metabolic enzymes.

Possible Risks

Organisms Used

E. coli DH5alpha, Bacillus subtilis 168, Synechococcus elongatus PCC 7942, Spirulina spp.

Risks of Selected Organisms

We use two kinds of algae: Synechococcus elongatus PCC 7942 (non-toxic lab strain, recognized as a safe organism and exists in nature) and Spirulina spp (edible food supplement), and they will not pose a threat even if it escapes from the lab.

Organisms Used As Chasses

Escherichia coli: E.coli DH5 alpha
Bacillus subtilis

Risks of Chassis

Bacillus subtilis 168 (Generally Regarded As Safe) and E.coli strain are recognized as Biosafety Level 1 by US public Health Service Guidelines and pose less risk to humans, which are not known to consistently cause disease in immunocompetent adult humans, and present a minimal potential hazard to laboratory personnel and the environment.

Parts Used and the Risk

Basic parts information

Part: BBa_K2932000

Name: Carbonic anhydrase / pSB1C3
Natural function of part: an enzyme catalyzing interconversion between carbon dioxide/water and bicarbonate/hydrogen ions
Source: gDNA of Synechococcus elongatus PCC7942 or a plasmid got from Addgene
Usage: to increase CO2 dissolving rate
Species: Synechococcus elongatus PCC7942
Risk Group: 1
Disease: No

Experiment Design

We set up a system containing algae, enzyme and culture media. We prepare the culture media with standard media of BG-11 or baking soda, liquid fertilizer, leaf fertilizer to adjust the ratio of composition and change the illuminance, temperature to find out the appropriate environment to optimize algal growth. We prepare raw lysates from Bacillus subtilis 168 producing CA or CYP2E1 by breaking down by 0.1 um glass beads, we didn’t further purify the specific enzymes and just used the total lysates as active biocatalysts. To test CO2 elimination by CA, we use indoor CO2 (500~1000 ppm) or CO2 tanks (2000 ppm and 100%) to provide the gases and analyze CO2 concentration with infrared CO2 sensor in a chemical hood. As to test chloroform, we asked NCTU-Formosa team to help in a specialized chemistry and engineering lab.

Risks of the Experiment

Our experiment only compares the growth of algae under different conditions and the solubility of carbon dioxide, so it will not pose any danger. And for chloroform analysis, we collaborated with NCTU-Formosa team for help by experts and a specialized lab.

Project Usage

In a consumer product that ordinary people buy
In a small enclosed device

Risks of Usage

The device we designed only contains two organisms and two kinds of enzyme currently: algae (edible Spirulina spp and lab strain Synechococcus elongatus PCC 7942) and enzymes (human carbonic anhydrase and rabbit CYP2E1). Even if the device is unfortunately broken, the former won't pose a threat to the environment. While the latter will be degraded naturally. Since the materials we use won't have any negative effect, safety, security or ethical risks won’t be considered.

Risks Management

Biosafety Level 1

Expert Assistance

Dr.Pei-Hong Chen will oversee us when we are doing the experiments. He had a PhD degree in Molecular Biology and is familiar with synthetic biology experiments. Also,he have instructed our school's iGEM teams for years.

Guidnace of Risk Management

We have lab skills and safety training by our instructor, Dr, Pei-Hong Chen, before working in the biotech laboratory. We always strictly followed the guidance of biological laboratory regulation at Taiwan given by Academia Sinica, the national academy of Taiwan. See the guide book here.

Topics Learned

Lab access and rules (including appropriate clothing, eating and drinking, etc.
Responsible individuals (such as lab or departmental specialist or institutional biosafety officer)
Differences between biosafety levels
Biosafety equipment (such as biosafety cabinets)
Good microbial technique (such as lab practices)
Disinfection and sterilization
Emergency procedures
Transport rules
Physical biosecurity
Personnel biosecurity
Dual-use and experiments of concern
Data biosecurity
Chemicals, fire and electrical safety

Work Areas

Open bench
Biosafety cabinet

Risk Management Tools

Accident reporting (measures to record any accidents)
Personal Protective Equipment (including lab coats, gloves, eye protection, etc)

Team:Mingdao/Safety

Safety Regulations