Lab Experience Personal reflection – Isabella Chiardi
In September 2021, I was given the opportunity to do my first lab rotation in the laboratory of cancer experimental therapeutics run by Professor Carlo Stella.
The lab is focused on Next Generation Sequencing based on circulating cell free DNA (cfDNA) in classical Hodgkin lymphoma patients. During my experience, I was followed by Doctor Martina Di Trani as a tutor, who introduced me into a variety of different protocols and procedures. Their research aims at investigating the prevalence of gene mutations in a large cohort of cHL patients, but also correlating the mutational level of cHL to the outcome and identifying potential biomarkers of Minimal Residual Disease in cHL.
To this purpose, I was taught about all the protocols that are based on a specific workflow in order to identify the previously mentioned mutations. The latter involves the following procedures: blood sample collection and processing, DNA extraction and quantification, library preparation and its quality check, target deep sequencing and final somatic analysis of the different mutations. Therefore, I particularly learned how to collect plasma and peripheral blood mononuclear cells (PBMC), how to extract DNA by using the MaxwellR RSC Blood DNA Kit, how to quantify this DNA through the Qubit protocol or the Picogreen dsDNA Assay Kit method, and how to prepare the library of my DNA by using the KAPA hyper Prep Kit (ROCHE) with all its various steps, that include: end repair and A-tailing of the DNA, adapter ligation and amplification of the library through PCR.
I enjoyed this experience very much since it gave me a first insight of the research world and most importantly it taught me how to behave in a lab, how to correctly use the different materials and machines, and how to diligently follow the different protocols. Even though it lasted for one month only, I truly learned a lot: while at the beginning I had no idea about how the lab life worked, now I can tell that I have acquired some skills and knowledge on how to perform multiple procedures even on my own.
Indeed, I am looking forward to my next lab experience with the second laboratory rotation to further deepen my knowledge into the research field.
