Safety has always been a crucial matter for iGEM. Safety awareness needs to be accurately applied in practical experiments. Before officially starting the experimental investigation, our team had already received safety education from Professor Li online, which gave us a preliminary understanding of the laboratory's safety regulations and rules. During the offline training session, our mentor also introduced us to the correct use of experimental equipment and emergency measures for accidents.

 

1. It is essential to wear shoes in the lab.

2. Wear white work clothes when entering the laboratory, and wear corresponding gloves and protective equipment when doing experiments.

3. It is forbidden to drink or eat in the laboratory. Don't bring food utensils into the laboratory to prevent poison pollution. Don't drink chemicals. Wash your hands before leaving the laboratory and eating.

4. Use volatile chemicals and drugs in the fume hood.

5. In treating laboratory waste liquid, attention should be paid to classification and timely treatment. A large amount of waste liquid can be stored at different times, and the waste liquid should be prevented from mixing.

6. When using the medicine, everyone should use it under the coach's instruction. During the test, the range should be manageable, causing the liquid to splash and spray so that it can be thrown onto himself or others.

7. Do not try to taste the chemicals and use equipment which you are not familiar with.

1. The centrifuge must be balanced before use, the lid must be closed, and the required speed and time must be set afterward.

2. Set the pipetting volume: the standard adjustment method is to adjust from an extensive range to a small degree and rotate the scale counterclockwise. When changing from a small field to a vast range, it should be adjusted to exceed the set volume scale first and then back to the set volume to ensure the pipette's accuracy.

3. Assemble the liquid transfer gun head: Insert the liquid transfer gun vertically into the suction head, rotate it half a turn left and right, and tighten it.

(Attention: hitting the suction head with the pipette is undesirable. The long-term operation will cause the parts of the pipette to loosen because of impact, which will cause the knob for adjusting the scale to get stuck.)

4. Suction and discharge of liquid: Vertical aspiration:

The tip of the suction head is immersed below 3mm into the liquid level, and the pipettor head is pre-moistened and washed in the liquid 2 ~ 3 times. Before suctioning, it is essential to ensure the accuracy and precision of liquid transfer. (Attention: slow suction and slow release of the liquid.)

5. The liquid transfer pipettor with liquid suction should not be placed horizontally.

6. Do not use a large range pipettor to transfer a small volume of liquid.

7. The scale of the pipettor should be adjusted to the maximum after each test.

 

All strains, reagents, and consumables used in the experiment are safe, do no harm to the ecological environment, and do not cause any damage to our human body under the standard operation of our laboratory personnel.

In our experiments, the strains we only used are E. coli DH5-α and Pichia pastoris Yeast GS115, both of which require BSL-1 condition, the lowest biosafety level. Other genetic materials including primers were ordered from company.

 

All gene editing techniques used in our research are for scientific experimental research. We did not use any animal or human genes in our experiments.