Safety
General lab safety guidelines:

(1)Wear lab coats, gloves, and goggles if necessary.

(2)Snacks, beverages and other foods are forbidden to be eaten in the laboratory.

(3)Wear gloves before touching the experimental equipment.

(4)Experimenters with long hair should tie up their hair when operating the experimental equipment.

(5)Wear shoes with the toe closed.

(6)It is strictly forbidden to play in the laboratory.

(7)The biological waste generated by the experiment should be placed in a special container.

(8)Experimental equipment should be sterilized and placed in the laboratory after use.

(9)When operating the experimental equipment, you should be familiar with its operation process in advance.

Operation of clean bench

(1) The window should be closed before UV light is turned on/UV light should be turned off before the window is opened.

(2) Turn on UV light 20-30 minutes before use.

(3) The window should be open at any time the fan is on.

(4) The window opening should be just large enough for hands to operate on the bench.

(5) Sanitize hands and other equipment thoroughly before operating on the bench.

(6) Sanitized hands and other equipment should dry on the bench before lighting alcohol lamp.

(7) Alcohol lamp cap should be placed on the lamp, lifted, and placed on top again to turn off. fire.

(8) Turn off the light, fan, and close the window after use.

Operation of centrifuge

(1) Test tubes should be shut tightly before placing into centrifuge.

(2) Test tubes should be placed opposite of each other, and should contain even amounts of liquid for even centrifugation.

(3) Centrifuge cap should be shut tightly before operation.

(4) Watch the centrifuge reach the desired RPM before leaving to look for any malfunction.

Operation of Ultrasonic Cell Disruptor

(1) Ensure the machine is placed in a clean, dry area, free from moisture, extreme temperatures, dust, and corrosive gases.

(2) Prepare the samples appropriately, adding them to the appropriate tubes or containers with the necessary buffers or solutions.

(3) Inspect the transducer, horn, and other components for proper installation and condition.

(4) Adjust the frequency, amplitude, and sonication time based on the sample type and desired disruption level.

(5) Use short pulses with adequate pauses to prevent overheating and excessive degradation of the sample.

(6) Monitor and adjust the temperature during sonication, using ice baths or refrigerated systems if necessary.

(7) Never operate the machine without the horn immersed in a liquid sample.

(8) Avoid touching the horn tip to the container walls to prevent damage.

(9) Wear protective eyewear and gloves to minimize the risk of splashes or contamination.

(10) Observe the sample during sonication for signs of overheating, cavitation, or other undesirable effects.

(11) Adjust the parameters or stop the sonication if necessary to prevent damage to the sample or equipment.

(12) Carefully remove the samples and clean the tubes, horns, and any other exposed parts.

(13) Document the experimental conditions, observations, and outcomes for future reference.

(14) Turn off the machine and disconnect it from the power source when not in use.

(15) Regularly inspect and maintain the equipment according to the manufacturer's instructions.

(16) Replace worn or damaged parts promptly to ensure optimal performance and safety.

(17) Keep detailed records of each experiment, including the settings used, sample details, and any issues encountered.

Operation of High-Pressure Steam Sterilizer

(1) Check Water Level: Ensure the outer chamber of the sterilizer has the appropriate amount of water. Avoid operating with insufficient or excessive water levels.

(2) Place Items Correctly: Arrange the items to be sterilized loosely in the inner chamber to allow steam to penetrate effectively. Avoid overcrowding or tightly packing the items.

(3) Secure the Lid: Ensure the lid is securely locked in place before starting the sterilization process.

(4) Purge Air: Prior to sterilization, purge the air from the chamber by opening the exhaust valve until all air is expelled. This ensures even temperature distribution and effective sterilization.

(5) Check Pressure and Temperature: Monitor the pressure and temperature gauges to ensure they reach the required levels for sterilization. Adjust the heat source as needed to maintain the desired conditions.

(6) Automatic Controls: If the sterilizer is equipped with automatic controls, ensure they are set correctly and functioning properly.

(7) Cool Down: Allow the chamber to cool down naturally before opening the lid. Do not forcibly release the pressure as this can cause boiling over or container damage.

(8) Handle Carefully: When removing sterilized items, use caution as they may be hot. Wear protective gloves if necessary.

(9) Regular Cleaning: Clean the sterilizer and its components regularly to prevent the build-up of dirt, residue, or bacteria.

(10) Inspect and Replace Parts: Regularly inspect and replace worn or damaged parts such as gaskets, valves, and filters to ensure proper functioning.

(11) Electrical Safety: Ensure the sterilizer is properly grounded and that electrical connections are secure. Avoid using damaged power cords or plugs.

(12) Overheating Prevention: Monitor the temperature to prevent overheating, which can damage the sterilizer or cause a fire hazard.

(13) Personal Protective Equipment (PPE): Wear appropriate PPE such as gloves, aprons, and eye protection when handling hot items or chemicals.

(14) Always refer to the manufacturer's operating manual and instructions for specific guidance on using the high-pressure steam sterilizer.

Operation of PCR Amplifier

(1) Ensure Cleanliness: Ensure that the laboratory workspace, PCR tubes, and reagents are clean and free from contaminants. Use clean pipette tips and avoid cross-contamination.

(2) Program Verification: Verify the PCR program (including temperature settings, cycle times, and ramp rates) with the protocol or recommended conditions for your specific assay.

(3) Ensure Cleanliness: Ensure that the laboratory workspace, PCR tubes, and reagents are clean and free from contaminants. Use clean pipette tips and avoid cross-contamination.

(4) Program Verification: Verify the PCR program (including temperature settings, cycle times, and ramp rates) with the protocol or recommended conditions for your specific assay.

(5) Uniform Loading: Load PCR tubes uniformly in the block to ensure even heating and cooling during the reaction.

(6) Secure Lids: Ensure that tube lids are securely closed to prevent evaporation and contamination.

(7) Temperature Calibration: Regularly check and calibrate the temperature sensors to ensure accurate temperature control.

(8) Avoid Overheating: Monitor the PCR process to prevent overheating, which can damage the instrument or affect the reaction.

(9) Containment: Handle PCR products carefully to avoid cross-contamination with other samples or the environment.

(10) Safe Disposal: Dispose of waste materials, including used PCR tubes and reagents, according to your laboratory's waste management protocols.

(11) Regular Cleaning: Clean the PCR block and instrument surfaces regularly to remove residue and reduce the risk of contamination.

(12) Service and Calibration: Schedule regular service and calibrations for the PCR instrument to ensure it is functioning properly.

(13) Wear Protective Equipment: Wear appropriate protective equipment, such as gloves, lab coats, and eye protection, when handling samples and reagents.

(14) Containment Facilities: Work in a designated biosafety cabinet or containment area when handling potentially hazardous materials.

(15) Power Supply: Ensure the PCR instrument is connected to a stable and appropriate power supply.

(16) Grounding: Verify that the instrument is properly grounded to prevent electrical hazards.

(17) Document Procedures: Document all PCR procedures, including sample details, reagent volumes, and PCR conditions, for future reference and quality control.

Microorganism safety:

Laboratories should use biological safety cabinets, containment equipment and negative pressure air systems to create physical barriers. All wastes must be properly sterilized before disposal. Access to laboratory areas should be restricted and controlled.

Ethical Considerations:

We used only Escherichia coli DH5α and Saccharomyces cerevisiae 1974 strains as chassis organisms for our experiments. And our experiments do not involve any human and animal organs, genes or tests. All genetic modifications are performed on a limited basis for scientific research purposes.