Today’s experiment: RNA extraction of Day 3 mouse 1st molars
For those of you who are like “????”, I’ll explain.
Unfortunately, to effectively explain what today’s experiment is all about would require an in-depth explanation of my project up to this point as well as the whole history of molecular biology, neither of which I’m willing to do here. More will be explained in future posts, probably.
In short, I’m dissecting just the molars of 3-day-old mouse teeth and isolating their RNA, which will later tell me how much of a certain set of genes the cells in these teeth are making. I will be doing the same for 6- and 9-day-old mice, and comparing the gene expression between these 3 ages.
So this morning the grad student in the lab next door graciously agreed to let me use her mice, who had conveniently given birth to pups 3 days ago.
I do not like dissections, so I was planning on using these pups later so I wouldn’t have to dissect today, but then I decided to take on this project and I needed an experiment. So I just took 4 of those 3-day-old pups, expecting to have 4 nice samples of RNA by the end of the day, one from each pup.
If only it was that simple.
I’ll try to spare you all the gory details, but in short 3-day-old pups are TINY and I couldn’t for the life of me find their molars even with the aid of a microscope. But bless the soul of my fellow grad student who sat there with me and helped me poke around, but even then out of the 4 first molars of each mouse we were only able to find 2 per mouse. I inadvertently crushed or lost the rest.
Conclusion for the dissection: I need practice. Except dissections are those things you want to get once and never have to do again. Alas.
So then I did my RNA extraction experiment, and even though RNA is super touchy and will spontaneously break down if you breathe on it or don’t spray everything with “RNase-Away” every 30 seconds, I managed to get some RNA out of my samples, which is what the graph above tells me. It’s a pretty smooth graph which means I did a pretty good job there 🙂 The bottom right corner says the RNA concentration (aka how much RNA I’ve got in my tube), and ideally I’d like it over 200 ng/uL or so but 99.9 is the best I got out of my 4 samples. The other ones were between 30-60.
Well, it looks like I can use the 99.9 ng/uL in the next step, so it’s not a total loss. But I’ll be doing lots of dissections and trying this again in the next couple of days.
Did you get your dose of science for today? I sure did.
Many mice will be harmed in the making of this project, I’m afraid. Such is science.