TRAINING

As high school students with no prior laboratory research experience, we folowed a training program at the Institute of Biochemistry of the Romanian Academy. Our training was designed to prepare us for conducting the wet lab work for our project, while ensuring we observed the standards of laboratory safety and sterile technique.

Our initial training focused on laboratory safety protocols. We learned the importance of maintaining a sterile working environment through proper use of personal protective equipment, including lab coats and clean gloves, to protect both us and our experiments from contamination.

We received training on the proper use, maintenance and sterilization procedure of Biological Safety Cabinets and Laminar Flow Hoods. Proper handling and disposal of used tubes and agar plates was also discussed. We learned that an organized working space was critical for preventing cross-contamination between samples and for maintaining the integrity of our experimental results. We also became familiar with essential laboratory equipment including heat blocks and centrifuges, learning their proper operation before beginning our experiments.

Mastering sterile technique while handling bacterial cultures presented the greatest initial challenge. During our first attempts, we occasionally forgot to maintain sterility or to change pipette tips between samples. However, through practice and guidance from experienced lab personnel, these procedures gradually became routine. We became proficient with standard micropipettes of various sizes and volumes. The importance of using fresh pipette tips for each sample became clear as we understood how easily cross-contamination could occur, so immediately disposing of used tips became part of our workflow.

In order to maintain sample integrity, we learned to minimize exposure time when working with agar plates and to ensure microcentrifuge tubes were promptly closed after use.

Our training in bacterial transformation procedures represented was the next step in our training. We learned to prepare competent cells, understanding that this step was essential for making bacterial cell membranes more permeable to foreign DNA. The addition of plasmids carrying antibiotic resistance genes required precision and careful technique. Through repeated practice, we became comfortable working with multiple antibiotic resistance markers. We were also trained to execute the heat shock procedure at 42°C , using stopwatches to ensure precise timing.

The plating process on antibiotic-containing agar plates represented a more compicated step in our training. We used overnight incubation at 37°C and the appearance of antibiotic-resistant bacterial colonies the following day served as visual confirmation of successful transformation. We learned the importance of rapid but careful pipetting when transferring bacterial samples to agar plates, because delays could affect sample viability, and the correct use of sterile spreader bars, to maintain sterility while distributing bacteria across agar plate surfaces.

Our laboratory training was essential for the experiments we needed to execute for our iGEM project. The guidance and support received from the team of researchers at the Institute of Biochemistry made our training effective and rewarding. We enjoyed working in the lab and are very grateful for having had the opportunity to train alongisde passionate and knowledgeable researchers.