Well, glory hallelujah, because I think my stuff finally worked.
Yesterday was a gloriously productive day. A few days earlier, I'd tested my hypothesis about why I was getting such poor sequencing results (answer: I'd been reusing something I wasn't supposed to reuse), but my results were still a little lackluster. Then I found out that our lab manager's assessment of our polymer was incorrect. It had gone bad, so my lukewarm results might not have been my fault. I ran them again yesterday, and they had markedly improved...not a single well was blank, as opposed to my worst day (April 18), when almost half of them were.
Is my bad streak over? I hope so.
I also did some highly successful PCR yesterday. I gauge it as successful because:
a) my positive control worked, which means that the TBE buffer I concocted on Monday (for the first time) is good, and
b) half the wells had no results.
This is actually what I was looking for, because it was an entire plate of negative controls. One of my six PCR reagents is contaminated with DNA, and I've been trying to isolate which one. In June, I determined it wasn't the water or the master mix. Yesterday, the wells with new BSA still showed DNA, but the wells with the new Taq did not. It was my Taq polymerase. Of course, it had to be the most expensive reagent I work with, but at least now I know.
I have been in an unbelievably deep dark funk ever since Steve emailed me last week and told me he was concerned about my progress in the lab, especially because I only have funding for another year, and summer is supposed to be the big time to get this stuff out of the way. I have rarely felt like such garbage. Especially because my time out of the lab has been spent at two weddings, collecting samples on the coast, and making up about 10 weeks worth of work in the herbarium, which is the part of this job that actually pays me. Plus, I was having rotten sequence results, and my PCR was contaminated. Things were not good.
Not to mention I've been busy during the year getting A's in all my classes. I only need 10 more credits and I'm done.
But is the bad luck and black funk over now? I hope so. Steve is on vacation this week, and I would love to have something good to show him when he gets back.
I broke down crying when I had to leave home last week. I have never ever done that.
And discoving very good songs that are also very very sad don't help much either, thank you very much, Death Cab for Cutie's "I Will Follow You Into The Dark."