This Gel of the Day comes with a dedication to the unnamed UC Santa Cruz researcher whose car was blown up on Saturday by fruitcakes who want to liberate Drosophila.
The Fab Four in this case are four specimens of Diaperis maculata; John, Paul and George were collected together from a decomposing Piptoporus betulinus fruiting body. Ringo joined them later, having been recovered from a Ganoderma applanatum sporocarp. In each of the bosex, the first four lanes represent a 1:50 dilution and the second four a 1:100 dilution of the initial DNA solution. Lanes 1, 5, 9, 13, 17, 21, 25 and 29 are the nucLSU; 2, 6, 10, 14, 18, 22, 26 and 30 are nucSSU; 3, 7, 11, 15, 19, 23, 27 and 31 are cox1 amplified using the general Coleopteran PAT/JER primer set; 4, 8, 12, 16, 20, 24, 28 and 32 were an attempt to amplify cox1 using a custom-designed primer set that works on the other Tenebrionidae on which I've used it but apparently not on D. maculata. This is the same result I got last when I tried a competitive test of the cox1 primer sets on several species which also included another member of the same subfamily.
This might be a useful phylogenetic signal; it could very well indicate a mutation in the binding site for the new primer set which is conserved in all the other Tenebs for which I could find sequences (I could find none for D. maculata when I designed the primers). Moreover, an alignment from my previous attempt came up with an insertion and two transitions for D. maculata at a binding site that wasn't present in the other tenebs. I can't tell yet whether this is a species-specific situation or if the mutations are present in all members of the genus. I'm hoping to collect specimens of D. maculata' cousin, D. nigronotata, in Florida next month. If I can get sequences out of it, I will be able to say that much more about the evolution of genus Diaperis and perhaps about tribe Diaperini as well. That is, if Big Coleopterists don't Expel me over raising the slight possibility that the tribe might not be monophyletic and if no wackadoos with IEDs decide to set me on fire in my sleep.
Have I mentioned lately how cool it is to find something that nobody (as far as I can find in the literature) has seen before? This is the good stuff right here. This is why I get up in the morning and slave over a hot thermocycler.