14 April 2023
By: Ellie Malone and Ilhan Garad
Introduction
Over the past two weeks, we have been performing RNA isolation on anole (Anolis carolinensis) liver samples to prepare us for the brain sample we worked on this week. This RNA can be formed into cDNA, which we use in the lab to test our selected gene, HS6ST1. The RNA results from the concentration will be used to calculate the math needed for cDNA synthesis that we will use next fall semester in lab.
For the past two weeks we have been doing RNA isolations on liver samples of green anole lizards. This process requires homogenization of the tissue, incubation with chloroform, centrifuging, and several washes with buffers to collect pure RNA. At first, we started with one sample then too two samples. After RNA was isolated, we tested the purity and concentration of our sample by using a nanodrop. By pipetting a small drop of our sample on the platform, the machine tells us how concentrated the RNA is and we can also check the quality of our sample. Well also ran it on agarose gel to test its integrity. When the RNA is intact, it shows two bands on the gel, which you can see on our gel from 4/7/2023, with samples 37 and 40. These were both liver samples. This week, we performed an RNA isolation on our first brain sample by following the steps we did on the liver sample.
Conclusion
Throughout the week, we learned how to isolate RNA. We were able to work with new equipment like the homogenizer and chemicals such as chloroform, ethanol, buffer RW1 and buffer EPE. We also learned new techniques. As we worked on our RNAs, they all turned out good. They had good purity (~2.0), concentration, and showed the right band (clear band).
