Sample information

Caught near creek in wooded area. Caught in summer. Temperature was 79 degrees. It was found next to its carapice and is greyish-black.

Wings are long and narrow, widening towards the back of the arthropod. They are also very thin. They extend past the back of the arthropod and sit against the abdomen. Wings are translucent with strong green and yellow veins. The edges of the wings are veined, with especially thick veins on the bottom edges. It has 2 wings and 6 legs. The foreleg has a thick ovular upper part and a much thinner lower part of the leg. The hind legs has a thicker cylindrical upper part of the leg and thinner lower part that has small hairs. All the legs are bent inward. The head is flat with a rounded, triangular face. The mouth is in the center of the face and takes up most of it. It is ridged and roughly rectangular, but narrows away from the eyes. It has two eyes that are large and spherical. They are facing forward on either side of the face and protrude significantly. It has no antennae. The body has three regions and is roughly cylindrical with a smaller head. The abdomen is thickest closest to the thorax and the tapers off significantly. The top is mostly black with patches of brown. The bottom is brown and black. The legs are green and brown and the wing veins are green.

Methods

DNA extraction: we had to remove liquid from the tube in order to add buffers

PCR: went as planned

Gel electrophoresis, Arthropod CO1: gel #1 lanes – ladder, C-1, C-2, C-3, C-4, positive Wolbachia control, negative Wolbachia control, water control; gel #2 lanes – ladder, C-5, C-6, C-6 again, Wolbachia positive control, Wolbachia negative control, water control

We accidentally added C-6 DNA to two wells. All of the samples and the arthropod controls had DNA, but the water control did not.

Gel electrophoresis, Wolbachia 16S: gel #1 lanes – ladder, C-1, C-2, C-3, C-4, positive Wolbachia control, negative Wolbachia control, water control; gel #2 lanes – ladder, C-5, C-6 again, Wolbachia positive control, Wolbachia negative control, water control

We only added 3 microliters of W- on gel 2. Both W+ had Wolbachia DNA as did C-3

DNA sequencing: Sequence was pretty good. We saved all bases with Q scores > 30.

Results

All of the samples had a band for arthropod CO1 DNA. The Wolbachia positive control had a band for both arthropod and Wolbachia DNA. The Wolbachia negative control only had a band for arthropod DNA. The water control did not have DNA in any gel. This shows that we did not have any contamination.