6 ms·

I had a similar experience as a beginning grad student. A postdoc in the lab had developed a radioligand binding assay, which we were using to purify a small pr

I had a similar experience as a beginning grad student. A postdoc in the lab had developed a radioligand binding assay, which we were using to purify a small protein. And I could just not get it to work, no matter how careful I was. TL;DR - I was being too careful, and so too slow. Because the binding was transient. So once I focused on speed instead of precision, the assay worked fine.

dekhn 6y ago
I distinctly recall a situation where speed made a huge difference. I took an undergraduate lab at UCSC where you were given raw bacterial cells and had to use the original literature to purify restriction endonucleases with it. The course ran for a quarter, and on the first day the professor said: "Most of you are going to spend a few days planning, then running your purifications for a few weeks. If I were going to do this, I'd spend a couple weeks planning and then do the whole experiment in a day, because the moment you break the cells open, all the proteases will start eating up your restriction endonucleases and you will lose activity in a couple days. I've also found a few situations where I couldn't get somethign to work and it turned out the person who had it "working" wasn't running controls (positive or negative), so you didn't really know if it worked.