Learning and Advice by Kim!

I love being an intern and going to the lab every day. Everyone I've met has been amazingly understanding and friendly, especially my mentors and fellow interns. It has been a rough learning time for me and I've probably skipped more steps than the TUSD learning tiers would ever allow, but it has been quite the experience - like learning a new language through immersion. I have learned more about science in the past two weeks than a semester (or two ) of biology at school and enjoyed it a great deal more.

This week I got a little more responsibility and freedom with things like pouring and loading gels, making PBS, tris, and other buffers and reagents. I also saw some aspects of working in a lab that are less appealing like getting inspected by men from Risk Management and Safety, filling pipette tips, and adjusting pH. Unfortunately, they are necessary to producing good science. Filling pipettes is tedious, but if you can master the one-handed insert by strategically picking up two or three tips between your fingers the task becomes far more amusing and enjoyable. pH technique eludes me. I was so scared of adding too much of HCl that I took 'drop by drop' literally. In retrospect, I would say that is unnecessary, but patience is ultimately the only way to get through that one. Inspection is tough. My advice is to act busy and blend into the background because getting interrogated about personal glove removing habits is intimidating regardless of how simple it seems.

Now, on to science...

Aysen and I have continued doing protein and DNA purifications, ligations, and digestions, but they are no longer just procedures to me. We run at least three protocols at once, but the initial daze and confusion, the blind pipetting and such has subsided. Last week's focus for me was understanding individual steps of the protocol - after the buffer add what; why do we have a secondary antibody? I'm still learning more of every step, but now I'm stepping back - stepping back in a sense that I can look up at the bigger picture, the goals, the significance of result, where all those little steps add up to. We are studying the Enteropathogenic (EPEC) and Enterohemorrhagic (EHEC) strains of E. Coli, specifically the flagella of each. The flagella is the part of E. coli that allows the bacteria to be motile, to move up and down the colon for example. Does the flagella have an affinity to mucin? Specifically, does each monomer of the flagella have an affinity to the mucin - and further, does a section of each flagellin's DNA also have a reaction to mucin? Mucins are a component of mucus, secreted by the body as a line of defense against pathogens, such as EPEC and EHEC strains of E.coli. Our mucin samples were collected from a cow colon as EPEC and EHEC interfere with the digestive processes in this part of the body. We test each structure of the flagella with Western blots and immunofluorescence to determine affinity to mucin. This study not only focuses on flagella's role in immunity, but also components of the structure itself. We are digesting, purifying, and ligating plasmids to take up a DNA insert that codes for the flagella. After we have ascertained the plasmid has taken up the insert, we will send it to be sequenced and hopefully will find that the genes and the insert can be expressed properly.

This is science for science- yet another building block of Truth to be used by the community to come to far and reaching goals unbeknownst presently. By studying flagella, E.coli becomes more familiar to the scientific community. E.coli plays a crucial role in how lab science is carried out. So although it does not give us life-altering results immediately, it holds great potential to change our lives.

I'm excited to see what we'll accomplish next week!

P.S. Fellow interns, I second Mon-Ning's movie night idea.