Tuesday, March 29, 2016
Tales of a Bad Mother
You would think that after spending over a month creating and working with your own IPC mutant, you would know it really well. Know every detail about it – which calcium binding loop was a little off, if it was feeling a little negative sometimes, it’s plans for M1D6 – everything. Even though I felt I knew IPC mutant D23N really well, it’s ins and outs, affinity target columns very quickly caused me and my lab partner to question which IPC was really ours. When performing our purification, we put our labels on the column instead of the elution tubes, so when we disposed of our columns, the elution tubes were identical. Our samples were pure, we just didn’t know which tube held the wildtype (WT) IPC and which tube held our mutant IPC D23N. It kind of felt like being a mom who took a different baby home from the hospital.
We made a 50/50 chance guess as to which tube was which and continued as normal and figured we would find out when we got the fluorescence assay data back. I managed to put this ordeal out of my mind until we finally graphed our data. Thankfully, we were comparing our data to another groups wildtype and mutant and discovered that our wildtype was closer to their mutant and our mutant was closer to their wildtype. This was when the suspicion kicked in that we had ended up on the wrong end of our 50/50 chance. Ultimately, we took to the literature. Lucky for us, the Kd and n were listed for the WT IPC, and we concluded that one of our samples was far closer to those values. This was also the sample that matched more closely with the other team’s wildtype data, so we were pretty certain we had switched the labels at this point. It just goes to show, that even when things are going horribly wrong – like picked up the wrong kid from the hospital wrong – science is always better when supported by literature and teamwork.