By: Emma Friedenberg
This week in the Mattis Lab at Cedars-Sinai Board of Governors Regenerative Medicine Institute was filled with learning the technicalities and maneuvers of different procedures, equipment, and computer software. The most interesting and hands-on experience was RNA isolation, the purification of RNA from biological samples. The isolation of RNA with high quality is a crucial step required to perform various molecular biology experiments.
Ribonucleic acid is a polymeric substance present in living cells and many viruses, consisting of a long single-stranded chain of phosphate and ribose units with the nitrogen bases (adenine, guanine, cytosine, and uracil), which are bonded to the ribose sugar. RNA is used in all the steps of protein synthesis in all living cells and carries the genetic information for many viruses.
A rapid method of analysis for determining the identity of neural stem cells (NSCs) and their sublineages involves the early detection of differentiation markers tracked at the RNA level. This week, I learned the procedure of RNA isolation from cell lines. The first round of RNA extraction was performed on a small set of three cell pellets – one each for my two mentors and one for me. Each step was rotated between the three of us for learning practices. The next day, a total of 18 cell pellets of our cell line underwent RNA isolation which was executed by myself and my post-doctoral mentor. By the end of the procedure, 1 microliter of RNA was transferred to the computer analysis system. Fortunately, all of the cell lines achieved an RNA concentration above the contamination or error mark.