Not a particularly successful PCR run today. For some reason, the ribosomal small subunit DNA didn't get amplified. I suspect that either I may have used an incorrect primer by mistake or else one of the pair may have gone bad. It happens. Luckily, I still have plenty of stock DNA and can have another crack at it. At least I got the large subunit and one of the cox1 genes from all of the specimens (the last eight wells are just a negative control; it's a good thing there's nothing there). I got nucLSU DNA, so it must be an error on my part that explains why the nucSSU didn't show up.
Oddly, the last two wells were taken from a 1:100 dilution and smeared; the previous two bands from a 1:50 dilution prepared from the same specimen didn't. Smearing usually indicates an excess of product. I'm not sure why that would have happened in this case. It's a duplicate, anyhow, so that's not important, just a bit strange.
The first sixteen wells in this gel are local specimens, collected at Wachusett Mountain. The third set of eight are from one of the specimens I picked up during the MSA foray in Pennsylvania last week. I have three more of those to sequence, too, once I get everything straight with these three.
As for tomorrow's field work, any B. cornutus I find will be shipped off live to Louisiana State for investigation of fungal symbionts in their gut. I don't really need to sequence anymore of them for the time being. Hopefully, I'll get lucky and find species other than this one and Diaperis maculata tomorrow. Those two seem to be dominant around here, but I'm going to try looking in a couple of different niches and see what turns up.
The North Florida field trip is two weeks from tomorrow. I know there will be more diversity down there, but I'd still like to come up with a few more local tenebs before winter sets in.
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