As I’ve explained in my previous post, my main project this summer aims to quantify the changes in NRG1 mRNA expression in mice that have been infused with kainic acid to induce seizures compared to mice infused with a control PBS solution. While this topic is investigated using RNAscope technology, my days aren’t spent performing this one experiment for 8 hours non stop. In fact, I split my time amongst the multi day procedure RNAscope calls for as well as completing a number of different lab management tasks that support the work of a number of other ongoing projects.
The breeding core that maintains our mice lines provides us with tail clippings to confirm the genotypes of individual mice we work with. Once I pick up the tails, I check with my PI which of them are a priority to genotype, I digest the tails to extract the DNA which I then run PCRs for and finally image their gels. The tail digestion is about two hours of work total but split over two days and PCRs take half an hour to prepare, an hour and a half in the thermal cycler, and another half hour in the gel electrophoresis before imaging the final results. In the waiting times between all these steps, I bounce back to my main project of RNAscope and work on tasks like cryosectioning brains and mounting them on slides. Days that I run through RNAscopes go from 9am to 7pm since the procedure involves a lot of dying the brain slices with different amplification steps to make specific mRNAs fluoresce and baking the slides in between amplification steps. Finally imaging the now dyed slides occurs the following day and takes several hours since we are obtaining highly detailed pictures of the now fluorescing mRNA.
As I schedule out all these tasks day to day, there’s the occasional side project I get pulled into, time spent observing the mice work my coworkers do, and lots of analysis to do on my computer in my down time. I really enjoy bouncing around my different responsibilities as no two days are the same!
