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In order to enable our team members to conduct experiments safely and efficiently in the biological laboratory without being harmed by reagents and instruments, we have learned a lot of knowledge about safety.

We received instructions on biology lab safety online. The teacher introduced us to common experimental instruments such as PCR instruments, centrifuge, gel electrophoresis instruments, etc., and briefly introduced the use method and common mistakes to ensure that we used the instrument correctly. At the same time, the common wrong usage and harm of the experimental consumables provided by us are introduced.

In the offline laboratory, our experimental supervision teacher introduced experimental specifications, instruments, and reagents to us in a more practical way. When doing experiments, we must wear a laboratory coat embroidered with a name to prevent contamination; In order to protect our hands and prevent the COVID-19 epidemic, we must wear nitrile gloves and medical surgical masks and replace them regularly during the whole process. The relevant specifications for the use of pipettors, PCR instruments, ultra-clean benches, and centrifuges were strictly clarified to us to avoid contamination or explosion during use.

PCR interpretation

Introduction to centrifuge use

In the agarose gel electrophoresis experiment, we need to use the segments of nucleic acid dyes for dyeing in EtBr purposes, but learned before the experiment in the interpretation of EB is a kind of strong mutagen of causes cancer or birth defects, so we all the players in the experimental process strictly to wear gloves and masks, avoid contact with EB reagent in any form, At the end of the experiment, new gloves were replaced and EB waste liquid was treated before discharge to avoid pollution of organisms and the environment. We also took biological precautions when viewing the gel with ultraviolet light.

Our experiment involves two kinds of Escherichia coli chassis microorganisms, which use DH5α for biological replication of DNA and BL21 for protein expression. These two kinds of Escherichia coli are quite safe, non-toxic, and harmless, and the leakage of genes in the experimental process is also avoided to ensure the safety and pollution of the environment.

Explanation of EtBr nucleic acid dye

Considering human safety and gene leakage, we did not plan to use BL21 containing chitinase as the main ingredient of our product. Instead, we designed a cell wall cleavage mechanism that uses arabinose to activate SRRz cleavage genes, change cell membrane permeability, degrade peptidoglycan in the cell wall, and finally break the cell wall. To achieve the purpose of releasing the product chitinase outside the cell, after re-purification, the harmful or irrelevant substances are removed, and a high concentration of the enzyme is obtained as the product. This mechanism avoids the direct action of BL21 on the human body, avoiding the harm of possible gene leakage.