1 Why It Is Necessary To Make Sure The Lb Agar Solution Is Cool Enoug ✓ Solved

1. Why it is necessary to make sure the LB-agar solution is cool enough before adding the ampicillin? What would happened if you add antibiotic when the solution is too hot? 2. Why the word gently emphasized so often in the procedure for making competent cells and what is happening to the cells during transformation?

3. Why do we put the LB-agar plates after transformation upside down in an incubator? Executive Program Practical Connection Assignment At UC, it is a priority that students are provided with strong educational programs and courses that allow them to be servant-leaders in their disciplines and communities, linking research with practice and knowledge with ethical decision-making. This assignment is a written assignment where students will demonstrate how this course research has connected and put into practice within their own career. Assignment: Provide a reflection of at least 500 words (or 2 pages double spaced) of how the knowledge, skills, or theories of this course have been applied, or could be applied, in a practical manner to your current work environment.

If you are not currently working, share times when you have or could observe these theories and knowledge could be applied to an employment opportunity in your field of study. Requirements: Provide a 500 word (or 2 pages double spaced) minimum reflection. Use of proper APA formatting and citations. If supporting evidence from outside resources is used those must be properly cited. Share a personal connection that identifies specific knowledge and theories from this course.

Demonstrate a connection to your current work environment. If you are not employed, demonstrate a connection to your desired work environment. You should NOT, provide an overview of the assignments assigned in the course. The assignment asks that you reflect how the knowledge and skills obtained through meeting course objectives were applied or could be applied in the workplace. 

Paper for above instructions

1. Importance of Cooling LB-Agar Solution Before Adding Ampicillin

LB agar (Luria-Bertani agar) is a nutrient-rich medium commonly used for culturing bacteria. One critical aspect of preparing LB agar with antibiotics, such as ampicillin, is ensuring that the agar solution is cooled adequately before the antibiotic is added. If ampicillin is introduced to a hot LB agar solution, its efficacy may be compromised (Davis, 2006).
Ampicillin is a β-lactam antibiotic that inhibits bacterial cell wall synthesis; it needs to remain stable to exert its effects effectively (Moore & Witte, 2017). High temperatures can lead to the degradation of ampicillin, resulting in a lower concentration of the antibiotic in the final medium. This degradation occurs because enzymes and chemical reactions can be activated at elevated temperatures that would otherwise remain inactive at lower temperatures (Harris et al., 2010). If the antibiotic is degraded before it can affect the target bacteria, transformed cells can grow unimpeded on the medium, leading to inconsistency in experimental results and undermining the entire purpose of using antibiotic selection.

2. Emphasis on Gentle Handling During Competent Cell Preparation

The term "gently" is emphasized throughout the procedure for making competent cells because bacterial cells, primarily E. coli, are fragile during the transformation process (Zhang et al., 2015). Making competent cells typically involves manipulating the cells in such a way (e.g., using calcium chloride) that enhances their uptake of foreign DNA, but excessive force or agitation can cause damage to the cell membranes (Sambrook & Russell, 2001).
During the transformation, cells undergo several phenomena: they transition into a state that allows them to take in plasmids or DNA fragments. This transformation process involves making small holes in the cell membrane, which can result in cell lysis if. Gentle manipulation helps preserve cell integrity, ensuring that a maximum number of cells remain viable and capable of taking up plasmid DNA (Khan et al., 2017). Over-agitation can lead to fewer successful transformations and an increased cell death rate, compromising experimental data integrity.

3. Why We Incubate LB-Agar Plates Upside Down

After transformation, LB-agar plates are incubated upside down primarily to prevent condensation from forming on the lid, which could lead to contamination. If plates are incubated right-side up, moisture from condensation can drip onto the agar surface, creating an environment conducive to unwanted bacteria or fungi (Chuang et al., 2018).
Additionally, incubating upside down allows for better air circulation around the agar plates, enhancing the oxygen exchange crucial for the growth of aerobic bacteria like E. coli (Delaney, 2015). This approach improves colony morphology and overall growth pattern, essential for experiments requiring visible colonies after growth.

Application of Course Knowledge in the Work Environment

As a student in the field of biological sciences, my coursework has equipped me with vital skills that I can apply in my future career as a microbiologist. These skills include a deeper understanding of microbial growth, genetic transformation, and laboratory best practices. The practical lab assignments fostered critical thinking and emphasized the importance of meticulous methodologies when working with live organisms.
For instance, my experience handling LB agar and antibiotics directly translates to tasks I might encounter in microbiological research or medical labs, where the culture of bacteria is fundamental for testing antibiotic resistance or for producing recombinant proteins. Understanding the importance of pH, temperature, and nutrient composition directly affects the quality of experimental outcomes. As stated by Beck et al. (2020), laboratory techniques such as agar plate preparation and transformation methods serve as foundational skills in synthetic biology.
Moreover, through thriving in a lab environment, I learned the value of teamwork and collaboration. Biology-dependent projects often include multiple stakeholders—for example, when developing new biotechnologies or antibiotic testing protocols. Effective communication among lab partners ensures that objectives are clearly understood and enhances the collective ability to troubleshoot issues during complex experimental procedures (Rosenberg et al., 2021).
Additionally, the ethical considerations in laboratory practices emphasized during the course have prompted me to contemplate my responsibility as a scientist. This ingrained sense of ethical awareness will guide my decision-making in my future work. For example, in research, ensuring that my work adheres to ethical standards related to genetic manipulation would be paramount. Ethical decisions, such as those related to the use of genetically modified organisms (GMOs), require a balance between scientific advancement and societal implications, a perspective underscored throughout the coursework (Keller & Mamo, 2020).
Overall, as I progress towards a career in biological research, the principles of experimental design, critical analysis, and ethical responsibility learned in this course will profoundly shape my professional trajectory and the nature of my contributions to the field of microbiology (Nielsen et al., 2014).

References

1. Beck, J. T., M, A., & Zrimsek, D. (2020). Laboratory Protocols in Microbiology. Academic Press.
2. Chuang, T. S., Lin, H. H., & Yang, C. P. (2018). Overcoming condensation effects on agar plates by using inverted plates during incubation. Journal of Experimental Biology, 220(14), 2581-2584.
3. Davis, R. W. (2006). Laboratory Procedures in bacteriology: Prevention of antibiotic degradation. Current Protocols in Microbiology, 3(1), A.4F.1-A.4F.14.
4. Delaney, J. (2015). Aerobic growth of bacteria on agar plates: An experimental perspective. Applied Microbiology, 11(2), 121-128.
5. Harris, S. A., Baker, B. M., & Holt, S. T. (2010). Effects of heat on β-lactam antibiotics. Therapeutic Drug Monitoring, 32(4), 487-491.
6. Keller, E. F., & Mamo, L. (2020). The ethics of biotechnology: Gene editing and societal issues. Nature Reviews Genetics, 21(4), 253-267.
7. Khan, M. A., Ali, W., & Maqbool, N. (2017). Competent cell preparation: Rescue of E. coli from clonal failure. Asian Journal of Biotechnology, 22(4), 56-64.
8. Moore, M. & Witte, W. (2017). Antibiotics: A global perspective on efficacy and resistance. Springer.
9. Nielsen, A. J., Thomsen, K., & Gram, L. (2014). Advances in microbial biotechnology for the development of new therapeutics. Microbial Biotechnology, 7(3), 298-312.
10. Rosenberg, J., Zisook, S., & Rangel, B. I. (2021). The importance of teamwork in laboratory research: Perspectives from lab scientists. BMC Medical Education, 21(1), 150.
This extensive analysis shows how integral fundamental lab practices are when handling living systems and applying theoretical knowledge to real-world problems, ultimately reflecting on a holistic education experience.