Continuing manuscript madness ...

... so not a lot of blogging today. However, I will take this time to complain about individuals who do not proof-read their manuscripts. My lab is currently working on a project looking at the denitrification pathway in various agricultural settings. So we're doing a combination of Real Time PCR (to look at gene copy density) and DNA analysis (RFLP and sequencing) to try to tease out some relationships. Well, there are several steps in the process, and all of them (save the final step) involve at least two enzymes. So we have a lot of analysis to perform.

Well, for the RFLP (restriction fragment length polymorphism) work we need PCR products which are long enough to be worthwhile to restriction digest. As you can guess, the "universal" primers we use aren't really universal, so it's a mixed bag. Add to this the fact that one of the primers we ordered IS WRONG and kept giving us erroneous results (about 300bp shorter than what it should have been). Turns out, the primer I ordered was based on a manuscript where the authors tacked on two 3' T's to the end of the primer (without an explanation) while citing the original paper. Figuring I had the correct primer sequence (should be identical in both papers, natch), I ordered it and didn't check it against the original. Doh!

At any rate, that drama consumed part of my day yesterday and I hope to close my office door, open up my manuscript word document, and spend a majority of today writing ... because tomorrow is filled with meetings.