Archive for January 20th, 2006

Mango Taq

Friday, January 20th, 2006

News from the Vendor Show:

A big PFTTTTTTTTTTT! for this innovation. Add some orange color and sell your polymerase as Mango Taq! It leaves an A overhang! Note to BioLine’s QC team: it leaves an A overhang because it doesn’t have proofreading ability so pehaps you should market is as a new and improved ” low-fidelity Taq for all your mutagenic needs!”
Verdict: I’ll stick with Taq Plus Precision from Stratagene.
Rumor mill: a favorite sales rep changes jobs……..goodbye and thanks! See you at the bar!

Yeast Screens

Friday, January 20th, 2006

My current project is to try to map the NLS and the import factor of my favorite protein. The problem is: everything I have tried has failed and I’m no closer to my goal than I was at the beginning of December. With the holidays, I decided to set up a yeast 2-hybrid library screen and let it cook while I was out of town.

Well….when I got back to work, colonies had grown on the plates and the work was just beginning. I grew the colonies up, assayed them, and then rescued the library DNA. I then grew up these colonies and purified the DNA. After sending the DNA down to be sequenced and re-transforming it to verify the interaction….what do I have? Six proteins that specifically interact with my bait protein but none of which are import factors!

So here’s my question: where the heck do I go next? I’ve done blast searches and pulled up countless papers on these proteins. How do I know what’s relevant and what should get thrown in the garbage. I, of course, don’t want to admit defeat so I’m exploring a few of the possibilities.

The thing that’s really frustrating about all of this is that technically the library screen worked perfectly. I just didn’t get what I wanted.

What I have learned is this: LUCK plays a major role in science and I don’t have any.