For me, there are things that I do on a daily basis, but there are also things that I do that depend on how a previous day’s work turned out.
Most days, I arrive at the lab I check in with my mentor Brittany and plan out the day’s work. Then we usually check on our cells to see if we need to split them, how to split them (1:10, 1:20, etcetera), or if the cells just need a media change.
Once we make a plan I get to work. On a given day, I might be making gels, and running gels, to get ready to develop a western blot the next day. I might also be collecting cells, and lysing the cells to either do an Immunoprecipitation (IP) or a normal western blot. Additionally, I will sometimes treat the cells, or transfect the cells in order to delete a certain gene or express a construct so that I can see what is happening to expression levels when I treat my cells. Finally, I may also step away from the cells and attempt to do a mutagenesis reaction. In order to do this, I will first set up the mutagenesis reaction (it is similar to a PCR reaction but with a different primer), and run the mutagenesis reaction. The next day (day 2), I will do a DPN1 to get rid of the original template (so that you have your mutated DNA). Then I will transform the new DNA into bacteria, and plate the bacteria. The next day(day 3), I will collect various colonies and grow them out. Then on day 4, I will isolate the DNA from the bacteria and send it off for sequencing. On day 5, I will receive the sequence and go from there.
Some days, the science does not go as you hoped it will and some experiments have to be done again, some might have to be changed, or you might have to stop the experiment altogether. All in all, the days in the lab in my experience is slightly different each day and builds upon the previous day. Each day, I learn something new and it has been a great experience. I am learning to be patient and realize it is okay if things don’t go as well as you hoped… more on this on next weeks blog where I will talk about the highs and lows of my project.
