Lusi vs. Insatiable, Unattainable Standard
I just write to keep my mental health in check
I have generated a protein A and B stably expressing cell from certain cell line. All of this was achieved after blood, sweat, and tears. The big boss set a standard too high, that even little boss (who previously doing the same thing) didn't have to achieve that standard back then. Little boss even knows this standard is ridiculous and defended me, but as always, the big one wins. As I heard, little boss only did activity test checking without having to prove that his stable cell line (SCL) it's 100% expressing his protein of interest. Another Japanese labmate was entrusted to make the same thing in another cell line with different protein. The particular antibody she is using with doesn't work well, so instead of checking the protein another senpai advised her to do qPCR instead. Even then there's no result coming out of them and big boss just assume that she did well and already generated the cell line the big boss wanted. Just now, a few hours ago, I was looking through her qPCR result. There's no significant deltaCt and I don't even think they're using the right calibrator. qPCR in and of itself isn't a suitable method for checking protein expression. Knowing this result, I predicted that integration of gene of interest didn't even happen. How come you want to express something? I have worked myself out, searching protocol by myself, as I know I am indeed a fool and I don't want to ask about fool. I struggled through failures, where everyone else failed and leave, and knowing their protocol is flawed, I tried again. Yesterday I was asked by big boss to gather information of protocol for making SCL, as if I'm still failing and my protocol is wrong. While in reality I believed that I am the second person (after little boss) who succeed. Whose protocol are you supposed to follow? Failed one? So you are comparing strange qPCR result with fluorescence imaging result? And those weird Ct value wins??? If Ct value can assure the cells are 100% expressing the protein, I would design the primer at oncr and don't give an F to florescence assay. I worked there times the effort with tripled standard (compared to those other J people) he put on my shoulder. Before, I wouldn't dare saying this is a form of racism, but seeing how big boss handles his international student, dare I say how implicit is this racism taking action. If you can't believe the ability of someone else from abroad, then just don't accept any international students. No matter how many times y'all say 頑張って or 一所懸命, I'll just work like a horse chasing unattainable, insatiable standard.