This week I faced a little bioethics dilemma.
I needed to test my primer pairs before I ran my precious samples. It would be best to test them with freshly isolated RNA, since if they failed I would have more confidence it was the primer pairs that failed, not the fact that I had bad RNA. So I went up to the rat room, got a rat, sacrificed it and took out its spinal cord. I took the tissue back to the lab and started the RNA isolation.
Then I made a mistake. I have been using the RNeasy kit for lipid-rich tissues on my RNA isolations. I use Qiazol for the lysis buffer. Both RNeasy and Qiazol have protocol cards and I was looking at the Qiazol card instead of the RNeasy card. The moment after I did the first step on the Qiazol card that is different on the RNeasy card, I realized my error. But it was too late. I continued on with the Qiazol method.
Was my RNA any good?
I could proceed with the RT-PCR, which takes several hours and reagents that aren’t cheap. Or I could assume it was no good and go get another rat. This rat came out of the colony and I have it to spare, so the only consideration is that the life of the first rat would be wasted.
I’m sure many (most?) scientists would laugh at such a consideration. Grant money and limited reagents are far more valuable than a rat.
I went ahead with the experiment. Most of the primer pairs I was testing did work, so this experiment was a success.
But if it hadn’t worked, I’d just thrown away roughly 2 days. (Because some steps can require a long block of time or equipment that others are using, scheduling can be tricky, and it may end up taking more time than it might take otherwise, for those of you who are saying “What’s the big deal? An RT-PCR shouldn’t take days.”)
Of course, it dawned on my later that I should have run the RNA on a gel. I don’t like to do that because I’ve had good RNA that looked bad because it degraded on the gel. It’s time consuming enough that I don’t really think it’s worth it. However, it was probably worth one rat’s small life.
By the way, I felt no guilt at all for making the mistake in the first place. These things happen to even the most experienced and careful of scientists. It is wise to factor in the mistakes in the planning. We even factor it into the rats. We order 10% extra for unforeseen circumstances: mistakes, a rat dies unexpectedly, or a blizzard keeps us from getting into the building to do the experiment.