ing, centrifuging many buffers that serve different purposes, but in the end help to purify our plasmid product. After the plasmid is purified, the concentration of nucleic acid in the sample is measured using a Nanodrop machine. Once this is completed, the sample is ready to be sent off for sequencing. This sequencing allows us to identify into which sites the plasmid has incorporated our non canonical amino acid (ncAA) mutation. Once those sites are known and isolated, it allows us to plate plasmid containing bacterial colonies with a single site mutation at different temperatures. I must say, I love being in the lab and getting the privilege to play even a small part in this work because its broader implications could be massive, and the work is equally as fascinating as it is challenging.
The first photo below shows an apparatus that is hooked up to a vacuum where wash buffers are used for the plasmid purification. And the second picture shows the Nanodrop machine.
It has been a great week filled with learning and growing as a scientist! Even as a teacher, consider myself a lifelong learner. So, having the opportunity to strengthen my science skills and understanding has been tremendous!
In Dr. Xiao’s lab, We are working a project that is incorporating synthesized non-canonical amino acids (ncAA) (amino acids that are outside of the 20 that occur in nature) to enhance protein thermal stability.
So this week’s activities have included making various solutions, incubating, culturing and plating E.coli cells, sending plasmids samples out for sequencing, analyzing sequences of plasmids to look for incorporation of our mutant ncAA and comparing it to wild type amino acid sequences for the enzyme Beta Lactamase. There’s also many smaller steps in between, and in between each step, I am learning new procedures, calculations and mindsets.
And even though I have been teaching science for years, I had never done something as routine in a lab as using an autoclave. I learned how to operate one this week.
So all around, I’ve been learning lots, and look forward to learning even more in the weeks to come.
This is a picture of me plating a culture of e-coli cells. (And if you at all concerned about the open flame in the picture, don’t be! It’s to sterilize the ethanol soaked tool that we use to spread the bacteria culture onto the plate smoothly.)